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Expression of Nerve Growth Factor Receptors and Their Prognostic Value in Human Breast Cancer1

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Abstract

Nerve growth factor (NGF) has been shown recently to be mitogenic for human breast cancer cells. In the present study, we have assayed the expression of NGF receptors (NGFRs: TrkA and p75) mRNAs in 363 human primary breast cancers, using real-time quantitative reverse transcription-PCR. NGFRs were found in all of the tumor biopsies. TrkA and p75 were positively correlated and were respectively associated with the histoprognostic grading and the tumor type. NGFRs were both related to progesterone receptors. In univariate analyses, TrkA (>upper quartile) was associated with longer overall survival. Histoprognostic grading, tumor size, node involvement, and steroid receptors were also prognostic factors. In Cox multivariate analyses, TrkA was not a prognostic parameter. This study demonstrates the expression of NGFRs in breast cancer and points out that patients with high levels of TrkA have a more favorable overall survival prognosis.
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2001;61:4337-4340. Cancer Res
Simon Descamps, Valérie Pawlowski, Françoise Révillion, et al.
Prognostic Value in Human Breast Cancer
Expression of Nerve Growth Factor Receptors and Their
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[CANCER RESEARCH 61, 4337–4340, June 1, 2001]
Advances in Brief
Expression of Nerve Growth Factor Receptors and Their Prognostic Value in
Human Breast Cancer
1
Simon Descamps, Vale´rie Pawlowski, Franc¸oise Re´villion, Louis Hornez, Mohamed Hebbar, Be´noni Boilly,
Hubert Hondermarck, and Jean-Philippe Peyrat
2
Laboratoire d’Oncologie Mole´culaire Humaine, Centre Oscar Lambret, 59020 Lille [V. P., F. R., L. H., M. H., J-P. P.], and Equipe Facteurs de Croissance, Laboratoire de
Biologie du De´veloppement, UPRES-EA 1033 Universite´ des Sciences et Technologies de Lille, 59655 Villeneuve d’Ascq [S. D., B. B., H. H.], France
Abstract
Nerve growth factor (NGF) has been shown recently to be mitogenic for
human breast cancer cells. In the present study, we have assayed the
expression of NGF receptors (NGFRs: TrkA and p75) mRNAs in 363
human primary breast cancers, using real-time quantitative reverse tran-
scription-PCR. NGFRs were found in all of the tumor biopsies. TrkA and
p75 were positively correlated and were respectively associated with the
histoprognostic grading and the tumor type. NGFRs were both related to
progesterone receptors. In univariate analyses, TrkA (>upper quartile)
was associated with longer overall survival. Histoprognostic grading,
tumor size, node involvement, and steroid receptors were also prognostic
factors. In Cox multivariate analyses, TrkA was not a prognostic parame-
ter. This study demonstrates the expression of NGFRs in breast cancer
and points out that patients with high levels of TrkA have a more
favorable overall survival prognosis.
Introduction
NGF
3
is an essential neurotrophin involved in the control of sur-
vival and differentiation of central and peripheral neuronal cells of
both the sympathetic and the sensory nervous system (1). NGF inter-
acts with its target cells through two categories of membrane binding
sites. The high affinity receptor tyrosine kinase (p140
trkA
) corresponds
to the TrkA proto-oncogene, and a secondary receptor known as p75
neurotrophin receptor (p75) is a member of the tumor necrosis fac-
tor-
␣
receptor superfamily and has no tyrosine kinase activity. Al-
though it has been clearly established that p140
trkA
tyrosine kinase
activity leads to the stimulation of the mitogen-activated protein
kinase cascade, the signaling mediated by p75 remains controversial
(2). In addition to its neurotrophic function, several other activities of
NGF have been described, including chemotactism and stimulation of
proliferation. In human prostatic cells, NGF participates in tumor cell
growth and invasion (3, 4). This last effect is mediated by p140
trkA
and p75 and indicates that NGF is involved in prostatic carcinogene-
sis.
We and others have demonstrated recently that NGF is able to
stimulate the proliferation of breast cancer cell lines (5, 6). We have
evidenced, using specific antibodies, the presence of p140
trkA
and p75
in MCF-7 and MDA-MB-231 cells. In addition, Tagliabue et al. (6)
have shown that p140
trkA
cooperates with p185
Her-2
in activating
growth of breast cancer cells. Altogether, these data suggest the
implication of NGF in breast cancer development and progression. In
previous studies, we have demonstrated that, in breast cancer biopsies,
insulin-like growth factor-1 receptors, fibroblast growth factor-2 re-
ceptors as well as type I growth factor receptors are related to tumor
prognosis (7–9). These results have led us to quantify the expression
of TrkA and p75 mRNAs in a series of 363 breast cancer biopsies. We
have shown that mRNAs for TrkA and p75 are expressed in breast
cancer, and we have evaluated their prognostic significance.
Materials and Methods
Cell Culture. Cell culture reagents were purchased from BioWhittaker
(France) except insulin, which was obtained from Organon (St. Denis, France).
The breast cancer cell lines (MCF7, T47D, BT20, SKBR3, MDA-MB-231,
MDA-MB-453, and MDA-MB-468) were obtained from the American Type
Culture Collection and routinely grown as described previously (10) in mono-
layer cultures. The SY5Y neuroblastoma cell line was a kind gift of Dr. Luc
Bue´e (INSERM U422, Lille, France).
Patients and Tumors. This study includes 363 patients who underwent
surgery for primary breast cancer in the Center Oscar Lambret (the Anti-
Cancer Center of the North of France), between May 1989 and December
1991. Tumor specimens were solely adenocarcinomas. At the time of collec-
tion, fat was removed, and samples were divided in two parts. The first part
was submitted for histological studies and HPG, according to the method of
Contesso et al. (11). The other part of the sample was immediately frozen in
liquid nitrogen for receptor assays (7).
The population studied is described in Table 1. The median age of the
patients was 58 years (range, 26–90 years). In prognostic studies, the median
duration of follow-up of living patients was 79 months. The number of relapses
(all distant relapses) was 126, and the number of patients who died from
intercurrent diseases was 94.
Total RNA Isolation. Total RNA was isolated from tumor samples (40 mg
weight) using the RNeasy Mini kit (Qiagen, Courtaboeuf, France), as described
by Pawlowski et al. (9).
Production of TrkA, p75, and TBP Standards. The mRNA standards
were obtained after in vitro transcription of a TrkA, p75, or TBP fragment
cloned in pGEM-T Vector Systems (Promega, Charbonnie´res, France) as
described by Pawlowski et al. (9); the transcription was carried out using the
RiboMAX Large Scale RNA Production System T7 for p75 and TBP (a
component of the DNA-binding protein complex TFIID), and the SP6 Pro-
duction System for TrkA (Promega, Charbonnie´res, France).
TrkA, p75, and TBP PCR Primers and TaqMan Fluorogenic Probes.
An amplicon of 89 bp was used for TBP, as described already (9). The PCR
primers and the TaqMan fluorogenic probes were designed using the Primer
Express software program (Perkin-Elmer; Demo version 1.0). TrkA sequences
were: forward, 5⬘-CATCGTGAAGAGTGGTCTCCG-3⬘; reverse, 5⬘-GAGA-
GAGACTCCAGAGCGTTGAA-3⬘; and probe, 5⬘-AGGAGTGAAATGGAA-
GGCATCTGGCG-3⬘. p75 sequences were: forward, 5⬘-CCTACGGCTACT-
ACCAGGATGAG-3⬘; reverse, 5⬘-TGGCCTCGTCGGAATACG-3⬘; and probe,
5⬘-CTCGGGCCTCGTGTTCTCCTGC-3⬘. Amplicons of 102 and 147 bp were
obtained for TrkA and p75, respectively, corresponding to sequences located in
the extracellular domain of each protein. The TaqMan probe carried a 5⬘
6-carboxy-fluorescein reporter dye in the cases of TrkA and p75 and a 5⬘VIC
reporter dye in the case of TBP. Primers and probes for TrkA and p75 and
Received 4/9/01; accepted 4/17/01.
The costs of publication of this article were defrayed in part by the payment of page
charges. This article must therefore be hereby marked advertisement in accordance with
18 U.S.C. Section 1734 solely to indicate this fact.
1
Supported by the Comite´ du Nord de la Ligue Nationale Contre le Cancer (Lille,
France).
2
To whom requests for reprints should be addressed, at Laboratoire d’Oncologie
Mole´culaire Humaine, Centre Oscar Lambret, rue F. Combemale, B.P. 307, 59020 Lille,
France. Phone: 33-3-20-29-55-35; Fax: 33-3-20-29-55-35; E-mail: jp-peyrat@o-lambret.fr.
3
The abbreviations used are: NGF, nerve growth factor; NGFR, NGF receptor; ER,
estrogen receptor; HPG, histoprognostic grading; OS, overall survival; PgR, progesterone
receptor; RFS, relapse-free survival; TBP, TATA box binding protein.
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probe for TBP were from Eurogentec (Seraing, Belgium), whereas primers for
TBP were from Genset (France).
Reverse Transcription-PCR Conditions. The reverse transcription and
the PCR were performed in a one-step methodology as described by
Pawlowski et al. (9), with optimal MgCl
2
concentrations of 6 mMfor TrkA, 3
mMfor p75, and 5 mMfor TBP. A template-free control was included in each
experiment. The non-template controls, standard RNA dilutions, and tumor
samples were assayed in duplicate.
Analysis and Expression of the Real-Time Reverse Transcription-PCR
Data. The quantification of the starting quantity of mRNA in an unknown
sample was performed by preparing a standard curve using dilutions of a
known amount of the respective standard RNA, as detailed by Pawlowski et al.
(9). The level of TrkA and p75 mRNA expression was expressed as a ratio
between receptor expression (in copies/
␮
g of total RNA) and TBP expression
(in copies/
␮
g of total RNA) and was referred to as normalized expression.
Statistical Analyses. All of the statistical analyses were done using SPSS
(version 9.0.1). The relationships between qualitative variables were deter-
mined using the
␹
2
test (with Yates correction when necessary). Correlations
between parameters were assessed according to the Spearman nonparametric
test. Comparison between subpopulations (PgR positive or PgR negative) were
assessed according to the Mann-Whitney nonparametric test. OS and RFS were
studied by Kaplan-Meier method analysis. The comparison of curves was
carried out by the log-rank test. Cox’s proportional hazard regression method
(12) was used to assess the prognostic significance of parameters taken in
association.
Results
Distribution of NGFRs in a Nonselected Series of 363 Human
Primary Breast Cancers. The distribution of breast cancer biopsies
according to their TrkA and p75 mRNA expression is presented in
Fig. 1. The median concentration of TrkA was 0.16, ranging from
0.005 to 3.48; the lower quartile was 0.079, and the upper quartile was
0.30. The median concentration of p75 was 0.19, ranging from 0.002
to 7.94; the lower quartile was 0.075, and the upper quartile was 0.52.
All of the studied breast cancer cell lines, used as controls, expressed
NGFRs. TrkA expression (TrkA:TBP) ranged from 0.0013 (BT20) to
0.012 (MDA-MB-468); it was 0.002 in MCF7, T47D, and MDA-MB-
231 and 0.005 in SKBR3 and MDA-MB-453. p75 expression ranged
from 0.0077 (MDA-MB-468) to 6.66 (MCF-7); it was 0.01 in MDA-
Table 1 Prognostic factors for patients according to TrkA and p75 status
TrkA p75
⬍Upper quartile
a
(%) ⱖUpper quartile
a
(%) P
⬍Upper quartile
b
(%) ⱖUpper quartile
b
(%) P
Age
⬍50 16.9 6.4 NS
c
15.8 7.1 NS
c
ⱖ50 58.3 18.5 59 18.1
Node
Negative 37.8 11.4 NS 35.4 13 NS
Positive 37.2 13.6 39.7 11.9
HPG
I 6.5 4.7 0.034 7 4.5 NS
II 37.3 10.9 35.4 12.7
III 32 8.7 32.2 8.3
Tumor type
Ductular 51.5 17.1 NS 56.6 11.8 ⬍0.001
Lobular 8 3.3 7.3 3.9
Others 15.4 4.7 11 9.3
Tumor diameter
⬍2 cm 6.3 2 NS 7 1.5 NS
2–5 cm 47.3 19.4 49.1 18
⬎5 cm 20.8 4.3 18.3 6.1
ER
⬍10 fmol/mg 21.5 4.7 NS 19.7 5.7 NS
ⱖ10 fmol/mg 53.1 20.7 56 18.6
PgR
⬍10 fmol/mg 22.4 5 NS 22.1 5.2 NS
ⱖ10 fmol/mg 52.1 20.4 53.6 19.2
a
Upper quartile, TrkA:TBP ⫽0.30.
b
Upper quartile, p75:TBP ⫽0.52.
c
NS, statistically not significant.
Fig. 1. A, distribution of TrkA mRNA expression in 363 nonselected primary breast
cancers. B, distribution of p75 mRNA expression in 355 nonselected primary breast
cancers. Tumors are distributed according to the relative expression of TrkA or p75. The
TrkA or p75 expression is normalized to the TBP expression and is expressed as a ratio
(TrkA or p75 expression:TBP expression).
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MB-453 and MDA-MB-231, 0.125 in T47D, 0.30 in SKBR3, and 0.5
in BT20.
Relationships between NGFRs and Other Parameters. Table 1
shows that TrkA expression levels are related to the HPG (
␹
2
test),
and that p75 mRNA expression levels are related to tumor type, with
low levels being more frequently found in ductular type tumors. A
positive correlation was found between TrkA and p75 (P⬍0.001),
and a negative correlation was found between TrkA and HPG
(P⫽0.034; Spearman test). Additionally, in the global population, we
have found positive correlation between ER and PgR (P⬍0.001), ER
and age (P⬍0.001), PgR and age (P⫽0.019), node involvement and
tumor diameter (P⬍0.001), and tumor size and HPG (P⫽0.01). We
found negative correlations between HPG and ER (P⬍0.001) on one
hand and HPG and PgR (P⬍0.001) on the other hand, as well as
between ER and tumor diameter (P⬍0.001).
TrkA (P⫽0.037), as well as p75 (P⫽0.026), were expressed to
a higher level in PgR-positive tumors compared with PgR-negative
tumors (Mann-Whitney nonparametric test). TrkA (P⫽0.030) and
p75 (P⫽0.054) levels were also higher in HPG I than in HPG II or
HPG III tumors. TrkA was found to be more highly expressed in small
tumors (⬍2 cm) than in large tumors (P⫽0.088). Finally, p75 level
was higher in menopausal patients (⬎50 years) than in nonmeno-
pausal patients (P⫽0.019).
Prognostic Studies. For Cox univariate analyses (Table 2; Fig. 2),
in OS studies, TrkA was a prognostic parameter, with high concen-
trations being associated with a good prognosis. HPG, tumor diame-
ter, node involvement, PgR, and ER were also prognostic parameters.
In RFS studies, only tumor diameter and node involvement were
prognostic parameters. Fig. 2 shows OS curves for all of the popula-
tion according to the expression of TrkA (threshold: upper quartile).
For Cox multivariate analyses, in OS studies, TrkA was not a
prognostic factor, whereas node involvement, PgR, and HPG were
prognostic factors. In RFS, node involvement, HPG, and tumor
diameter were prognostic parameters.
Discussion
In the present study, NGFRs were found to be expressed in all of
the breast cancer cell lines and breast cancer biopsies that we have
studied. We have found by immunoprecipitation and Western blotting
experiments that the proteins corresponding to TrkA and p75 gene
expression, p140
trkA
and p75
NTR
, respectively, are present in breast
cancer cell lines and are activated by NGF (5). Thus, NGFR mRNA
expression in breast cancer cells is associated with the expression of
the respective proteins. In our study, TrkA and p75 mRNAs were
found to be correlated in breast cancer and to be expressed in equiv-
alent amounts because their median concentrations and upper and
lower quartiles are very similar. These results, and previous demon-
strations that NGF is mitogenic for breast cancer cell lines (5, 6),
strongly suggest an implication of NGF in breast cancer development.
The TrkA tyrosine kinase receptor is known to mediate the NGF
mitogenic effect by activating the Ras/Raf/mitogen-activated protein
kinase pathway. p75 has been shown to regulate TrkA activation by
NGF, and it could also have a role in the regulation of apoptosis by
activation of a specific pathway involving ceramides, c-Jun NH
2
-
terminal kinase, caspases, or nuclear factor-
␬
B (13). p75 has been
shown recently to be responsible of the antiapoptotic effect of NGF on
schwannoma cells (14) and could have a similar role in breast cancer
cells (15). Considering the described effects of NGF, we can hypoth-
esize that this growth factor is able to modify the equilibrium between
proliferation and apoptosis and to favor tumor growth.
In breast cancers, there was a wide range of variation in TrkA
(0.05–3.48) and p75 (0.002–7.94) mRNA expression (normalized to
TBP expression). We demonstrate that a first source of variation is the
tumor type, because TrkA level is lower in high-grade tumors and p75
expression is higher in ductular carcinomas. We found that high
concentrations of TrkA were related to high concentrations of PgR.
The mechanism by which TrkA is regulated in breast cancer has never
been described. The observed coexpression of TrkA and steroid
receptors could suggest a common regulator of these receptors; there-
fore, we can hypothesize that estradiol stimulates the expression of
TrkA transcripts, because it has been shown for transcripts of insulin-
like growth factor receptors (16). Two studies have reported the
presence of NGF in milk (17) and in capsules surrounding breast
implants (18), showing the presence of NGF in the mammary gland.
Thus, the variation of NGFR expression that we have observed in
breast cancer biopsies could be related to NGF regulation of its own
receptors.
We demonstrate that a high TrkA mRNA level is associated with a
good prognosis in OS univariate analyses, with a median duration of
follow-up of 6.5 years. The best TrkA threshold defined was 0.30
(normalized relative to TBP), corresponding to the upper quartile. In
contrast, in multiparameter Cox analyses, TrkA was not a prognostic
parameter; this was not unexpected, considering its relation with
HPG. It might be considered as paradoxical that a tumor containing a
high level of receptors for the growth factor NGF exhibits a better
prognosis than a tumor without. Such a relationship has already been
reported in neuroblastoma (19). Moreover, we have reported relation-
ships between tyrosine kinase receptors and prognosis in breast cancer
for insulin-like growth factor-I receptors (7), fibroblast growth fac-
tor-2 receptors (8), and type I growth factor receptors (9). We can
hypothesize that tumors with high levels of TrkA receptors have
retained some physiological control of growth, which could explain
the better prognosis.
In conclusion, the present study emphasizes the idea of the involve-
ment of NGF in human breast cancer growth and points out that
Table 2 Prognostic factors in Cox univariate analyses
The thresholds (upper quartiles) were 0.30 (TrkA:TBP ratio for TrkA and 0.52
(p75:TBP ratio) for p75.
OS RFS
PRR
a
PRR
TrkA 0.034 0.56 NS
p75 NS NS
ER (⬍10; ⱖ10 fmol/mg protein) 0.044 0.64 NS
PgR (⬍10; ⱖ10 fmol/mg protein) 0.0042 0.54 NS
Node involvement (0; ⬎0) 0.034 1.56 0.0088 1.61
Tumor diameter (⬍2; 2–5; ⬎5 cm) 0.0038 1.75 0.0022 1.66
HPG (I, II, III) 0.0008 1.84 NS
a
RR, risk ratio; NS, statistically not significant.
Fig. 2. OS in the whole population according to the presence of TrkA. Curve 1, patients
with high level of TrkA expression (TrkA:TBP, ⬎0.30); Curve 2, patients with low TrkA
expression (TrkA:TBP, ⬍0.30). A higher OS was found in TrkA-positive patients.
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patients with a high level of TrkA receptors have a better prognosis.
Then NGFRs are potential targets for new breast cancer therapies, and
the recent demonstration by Tagliabue et al. (6) that p140
trkA
coop-
erates with p185
Her-2
in activating the growth of breast cancer cells
suggests that blocking the NGFRs could improve the effect of anti-
erbB2 drugs.
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on June 2, 2013. © 2001 American Association for Cancer Research. cancerres.aacrjournals.org Downloaded from
... More recently, Kumar et al. [94] reported an overexpression of NGF in benign phyllodes, a rare fibro-epithelial breast tumor, demonstrating that this neurotrophin could play a role beyond the pathogenesis of malignant tumors. In addition to NGF, NGFR/p75NTR and TrkA receptors are also expressed in breast cancers cells, either at the transcriptional or protein level [83,84,89,95]. ...
... In particular, protein and immunological expression of NGF have been reported in the majority of human breast tumors [82], making it a broader diagnostic potential than ER or HER-2 [17]. Research has also shown that TrkA and NGFR/p75NTR receptors are overexpressed in most types of breast cancer compared to normal cells [87,89]. Moreover, Islam et al. [14] demonstrated that Russell's Viper Venom (RVV)-NGFa (an NGF isoform), labelled with Fluorescein Isothiocyanate (FITC), establishes strong binding to TrkA and NGFR/p75NTR receptors in breast cancer cells but not in non-cancerous cells. ...
... The prognostic value of NGF, TrkA and NGFR/p75NTR expression was evaluated in several studies [15][16][17]19,83,87,89,93,99]. ...
Expression and Signaling Pathways of Nerve Growth Factor (NGF) and Pro-NGF in Breast Cancer: A Systematic Review
Article
Full-text available
  • Oct 2022
  • Curr Oncol
Breast cancer represents the most common type of cancer and is the leading cause of death due to cancer among women. Thus, the prevention and early diagnosis of breast cancer is of primary urgency, as well as the development of new treatments able to improve its prognosis. Nerve Growth Factor (NGF) is a neurotrophic factor involved in the regulation of neuronal functions through the binding of the Tropomyosin receptor kinase A (TrkA) and the Nerve Growth Factor receptor or Pan-Neurotrophin Receptor 75 (NGFR/p75NTR). In addition, its precursor (pro-NGF) can extert biological activity by forming a trimeric complex with NGFR/p75NTR and sortilin, or by binding to TrkA receptors with low affinity. Several examples of in vitro and in vivo evidence show that NGF is both synthesized and released by breast cancer cells, and has mitogen, antiapoptotic and angiogenic effects on these cells through the activation of different signaling cascades that involve TrkA and NGFR/p75NTR receptors. Conversely, pro-NGF signaling has been related to breast cancer invasion and metastasis. Other studies suggested that NGF and its receptors could represent a good diagnostic and prognostic tool, as well as promising therapeutic targets for breast cancer. In this paper, we comprehensively summarize and systematically review the current experimental evidence on this topic. INPLASY ID: INPLASY2022100017.
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... More recently, Kumar et al. [92] reported an overexpression of NGF in benign phyllodes, a rare fibro-epithelial breast tumor, demonstrating that this neurotrophin could play a role not only in the pathogenesis of malignant tumors. Beyond NGF, also NGFR/p75NTR and TrkA receptors are expressed in breast cancers cells either at the transcriptional and protein level [93][94][95][96]. ...
... In particular, it has been reported a transcript, protein and immunological expression of NGF in the majority of human breast tumors [91], making it a broader diagnostic potential than ER or HER-2 [17]. Noteworthy, several studies have also shown the overexpression of TrkA and NGFR/p75NTR receptors in most of the human breast cancers as compared to the expression of these receptors in normal cells [94,114]. Moreover, Islam et al. [14] demonstrated that Russell's Viper Venom (RVV)-NGFa (an NGF isoform), labelled with Fluorescein Isothiocyanate (FITC), establishes strong binding to TrkA and NGFR/p75NTR receptors in breast cancer cells but not in non-cancerous cells. ...
... About NGF receptors, Descamps et al. [94] reported that the overall TrkA mRNA expression predicts a more favorable prognosis in a highly variable cohort of 363 primary breast carcinoma. However, as mentioned above, Davidson et al. [114] showed that the levels of p-TrkA were associated with tumor progression to effusion in metastatic breast carcinoma, thus correlating the dysregulation of p-TrkA with poor prognostic outcome in a more uniform cohort of 39 patients. ...
Expression and Signaling Pathways of Nerve Growth Factor (NGF) And Pro-NGF in Breast Cancer: A Systematic Review
Preprint
Full-text available
  • Sep 2022
Breast cancer represents the most frequent cancer and the leading cause of cancer death among women. Thus, the prevention and early diagnosis of breast cancer appears to be of primary urgency as well as the development of new treatments able to improve its prognosis. Nerve Growth Factor (NGF) is a neurotrophic factor that plays a key role in the regulation of neuronal functions thought the binding to the Tropomyosin receptor kinase A (TrkA) and the Nerve Growth Factor receptor or Pan-Neurotrophin Receptor 75 (NGFR/p75NTR). Also, its precursor (pro-NGF) can extert biological activity by forming a trimeric complex with NGFR/p75NTR and sortilin or by binding to TrkA receptors with low affinity. Both in vitro and in vivo studies showed that NGF is synthesized and released by breast cancer cells and has mitogen, antiapoptotic and angiogenic effects on these cells through the activation of different signaling cascades that involve TrkA and NGFR/p75NTR receptors. Conversely, pro-NGF signaling has been related to breast cancer invasion and metastasis. Other studies suggested that NGF and its receptors could represent a good diagnostic and prognostic tool, as well as promising therapeutic targets for breast cancer. Here, we comprehensively summarize and systematically review the current experimental evidence on this topic.
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... Les voies de signalisation sous-jacentes à l'axe NGF/TrkA ne sont pas tout à fait comprises dans le cancer du sein mais il a cependant été démontré que les voies de signalisation Pi3K/Akt et MAPKs, des voies retrouvées dans les cellules neuronales, sont activées pour promouvoir la croissance tumorale (Descamps et al., 1998(Descamps et al., , 2001b(Descamps et al., , 2001aDollé et al., 2003Dollé et al., , 2004. La protéine p21 WAF1/Cip1 serait également impliquée dans l'induction de la survie causée par p75 NTR dans les cellules cancéreuses mammaires (Verbeke et al., 2010). ...
... Le gène NTRK1 est majoritairement transcrit dans la lignée MDA-MB-231 tandis que la lignée BT-474 présente la plus faible proportion d'ARNs messagers pour ce gène (Figure 36.G et H). Par ailleurs et comme déjà reporté précédemment(Descamps et al., 2001a;Lagadec et al., 2009), le gène NTRK1 est faiblement transcrit dans l'ensemble des lignées cancéreuses mammaires car chaque lignée présente un Ct supérieur à 30 cycles. ...
... Pour TrkA et CD44 notamment, ces études ont souvent abouti à des résultats contradictoires. Ainsi, le laboratoire a démontré par analyse transcriptomique que l'expression de TrkA est liée avec l'augmentation de la survie(Descamps et al., 2001a) alors que Davidson et son équipe ont démontré que la phosphorylation de TrkA est un facteur péjoratif(Davidson et al., 2004). Pour CD44v3, il a été montré que les isoformes de CD44 contenant le variant 3 ne sont pas corrélées avec le pronostic(Wu et al., 2015) alors que Ryś et ses collègues montrent une association avec l'envahissement ganglionnaire(Ryś et al., 2003). ...
Plasticité des réseaux de récepteurs membranaires dans la signalisation du NGF et de son précurseur dans les cancers du sein
Thesis
  • May 2019
Le laboratoire INSERM U908 a montré le rôle déterminant du facteur de croissance NGF dans l’agressivité du cancer du sein. De précédentes études ont montré que l’inhibition de l’activité de TrkA, le récepteur du NGF, aboutissait à une diminution de la taille des tumeurs in vitro mais aussi dans des modèles pré-cliniques. Néanmoins, ces inhibiteurs ont donné lieu à des essais cliniques décevants. La résistance à ces inhibiteurs est en partie due à l’association de récepteurs membranaires. Plus précisément, mon travail de thèse a permis de démontrer qu’une forme particulière du récepteur CD44 interagit avec TrkA ce qui conduit à une résistance aux traitements. De plus, j’ai également montré l’existence d’une interaction entre TrkA et deux autres récepteurs. Ainsi, j’ai pu déterminer précisément comment ces récepteurs coopèrent afin de développer des inhibiteurs ciblant leur interaction. Ces inhibiteurs pourraient donc s’avérer efficaces pour le traitement de certains cancers du sein. Ma thèse souligne l’importance des complexes de récepteurs dans la plasticité des cellules cancéreuses et leur capacité à s’adapter pour résister aux thérapies ciblées.
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... These steps require dynamic interactions between cancer cells and the brain microenvironment [9]. In this context, we have shown that the tyrosine kinase receptor TrkA, was overexpressed in up to 20% [10] of TNBC cases and was involved in both tumorigenesis and the metastasis in vitro and in vivo [11][12][13][14]. TrkA is the high-affinity receptor of nerve growth factor (NGF) [15,16] and in TNBC, both NGF and its precursor the proNGF have been found to increase invasion and migration through TrkA-mediated mechanisms [12,17]. ...
... In this context, we have shown that the tyrosine kinase receptor TrkA, was overexpressed in up to 20% [10] of TNBC cases and was involved in both tumorigenesis and the metastasis in vitro and in vivo [11][12][13][14]. TrkA is the high-affinity receptor of nerve growth factor (NGF) [15,16] and in TNBC, both NGF and its precursor the proNGF have been found to increase invasion and migration through TrkA-mediated mechanisms [12,17]. These two growth factors act through TrkA phosphorylation and underlying signaling pathway [14,17]. ...
ProNGF promotes brain metastasis through TrkA/EphA2 induced Src activation in triple negative breast cancer cells
Article
Full-text available
  • Dec 2023
Background Triple-Negative Breast Cancer is particularly aggressive, and its metastasis to the brain has a significant psychological impact on patients' quality of life, in addition to reducing survival. The development of brain metastases is particularly harmful in triple-negative breast cancer (TNBC). To date, the mechanisms that induce brain metastasis in TNBC are poorly understood. Methods Using a human blood–brain barrier (BBB) in vitro model, an in vitro 3D organotypic extracellular matrix, an ex vivo mouse brain slices co-culture and in an in vivo xenograft experiment, key step of brain metastasis were recapitulated to study TNBC behaviors. Results In this study, we demonstrated for the first time the involvement of the precursor of Nerve Growth Factor (proNGF) in the development of brain metastasis. More importantly, our results showed that proNGF acts through TrkA independent of its phosphorylation to induce brain metastasis in TNBC. In addition, we found that proNGF induces BBB transmigration through the TrkA/EphA2 signaling complex. More importantly, our results showed that combinatorial inhibition of TrkA and EphA2 decreased TBNC brain metastasis in a preclinical model. Conclusions These disruptive findings provide new insights into the mechanisms underlying brain metastasis with proNGF as a driver of brain metastasis of TNBC and identify TrkA/EphA2 complex as a potential therapeutic target.
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... NGF and its receptors play a role in BC. TrkA levels have a prognostic value in BC patients (Descamps et al., 2001a), and secretory BCs are driven by oncogenic ETV6-NTRK3 gene fusions (Lee et al., 2014). NGF signaling fosters the survival and proliferation of BC cells (Descamps et al., 1998(Descamps et al., , 2001b, and the anti-estrogen tamoxifen inhibits this effect (Chiarenza et al., 2001). ...
... Many findings have highlighted the role of TrkA as a driver of cell transformation. They have also suggested that derangement of the NGF circuit is involved in drug resistance, survival, and metastatic spreading of solid tumors (Demir et al., 2016), including the so-called hormone-dependent cancers (Descamps et al., 1998(Descamps et al., , 2001aGeorge et al., 1998;Pflug and Djakiew, 1998;Sigala et al., 1999;Djakiew, 2001, 2002;Miknyoczki et al., 2002;Dollé et al., 2003Dollé et al., , 2004Festuccia et al., 2007;Papatsoris et al., 2007;Anagnostopoulou et al., 2013). Specific targeting of NGF inhibits the proliferation and metastatic events in BC (Adriaenssens et al., 2008). ...
Targeting the Nerve Growth Factor Signaling Impairs the Proliferative and Migratory Phenotype of Triple-Negative Breast Cancer Cells
Article
Full-text available
  • Jun 2021
Triple-negative breast cancer is a heterogeneous disease that still lacks specific therapeutic approaches. The identification of new biomarkers, predictive of the disease’s aggressiveness and pharmacological response, is a challenge for a more tailored approach in the clinical management of patients. Nerve growth factor, initially identified as a key factor for neuronal survival and differentiation, turned out to be a multifaceted molecule with pleiotropic effects in quite divergent cell types, including cancer cells. Many solid tumors exhibit derangements of the nerve growth factor and its receptors, including the tropomyosin receptor kinase A. This receptor is expressed in triple-negative breast cancer, although its role in the pathogenesis and aggressiveness of this disease is still under investigation. We now report that triple-negative breast cancer-derived MDA-MB-231 and MDA-MB-453 cells express appreciable levels of tropomyosin receptor kinase A and release a biologically active nerve growth factor. Activation of tropomyosin receptor kinase by nerve growth factor treatment positively affects the migration, invasion, and proliferation of triple-negative breast cancer cells. An increase in the size of triple-negative breast cancer cell spheroids is also detected. This latter effect might occur through the nerve growth factor-induced release of matrix metalloproteinase 9, which contributes to the reorganization of the extracellular matrix and cell invasiveness. The tropomyosin receptor kinase A inhibitor GW441756 reverses all these responses. Co-immunoprecipitation experiments in both cell lines show that nerve growth factor triggers the assembly of the TrkA/β1-integrin/FAK/Src complex, thereby activating several downstream effectors. GW441756 prevents the complex assembly induced by nerve growth factor as well as the activation of its dependent signaling. Pharmacological inhibition of the tyrosine kinases Src and FAK (focal adhesion kinase), together with the silencing of β1-integrin, shows that the tyrosine kinases impinge on both proliferation and motility, while β1-integrin is needed for motility induced by nerve growth factor in triple-negative breast cancer cells. The present data support the key role of the nerve growth factor/tropomyosin receptor kinase A pathway in triple-negative breast cancer and offer new hints in the diagnostic and therapeutic management of patients.
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... The mammalian cells expressing TrkA receptors such as PC-12, MCF-7, and MDM-MB-231 (0.5 × 10 5 ) (Descamps et al., 2001) were grown in 96 well cell culture plate in DMEM and then anti-TrkA, TrkB, TrkC or p 75 NTR primary antibody (1:1,000 in DMEM) or 1X PBS (control) was added to each well. The cells that did not express TrkA receptor such as HEK-293 and L6 were used as negative controls. ...
... In any case, similar to mouse 2.5S-NGF, Nn-α-elapitoxin-1 has also been found to be highly specific toward the TrkA receptor subtype of PC-12 cells. This study has provided conclusive evidence that Nn-α-elapitoxin-1 binds to TrkA receptor expressed on the surface of PC-12, MCF-7 and MDA-MB-231 (Descamps et al., 2001); however, the cells such as HEK-293 and L6 lacking this receptor on cell surface do not show binding with ...
Transcriptomic, proteomic, and biochemical analyses reveal a novel neuritogenesis mechanism of Naja naja venom α‐elapitoxin post binding to TrkA receptor of rat pheochromocytoma cells
Article
Full-text available
This is the first report showing unique neuritogenesis potency of Indian Cobra N. naja venom long‐chain α‐neurotoxin (Nn‐α‐elapitoxin‐1) exhibiting no sequence similarity to conventional nerve growth factor, by high‐affinity binding to its tyrosine kinase A (TrkA) receptor of rat pheochromocytoma (PC‐12) cells without requiring low‐affinity receptor p⁷⁵NTR. The binding residues between Nn‐α‐elapitoxin‐1 and mammalian TrkA receptor are predicted by in silico analysis. This binding results in a time‐dependent internalization of TrkA receptor into the cytoplasm of PC‐12 cells. The transcriptomic analysis has demonstrated the differential expression of 445 genes; 38 and 32 genes are up‐regulated and down‐regulated, respectively in the PC‐12 cells post‐treatment with Nn‐α‐elapitoxin‐1. Global proteomic analysis in concurrence with transcriptomic data has also demonstrated that in addition to expression of a large number of common intracellular proteins in the control and Nn‐α‐elapitoxin‐1‐treated PC‐12 cells, the latter cells also showed the expression of uniquely up‐regulated and down‐regulated intracellular proteins involved in diverse cellular functions. Altogether, the data from transcriptomics, proteomics, and inhibition of downstream signaling pathways by specific inhibitors, and the immunoblot analysis of major regulators of signaling pathways of neuritogenesis unambiguously demonstrate that, similar to mouse 2.5S‐nerve growth factor, the activation of mitogen activated protein kinase/extracellular signal‐regulated kinase is the major signaling pathway for neuritogenesis by Nn‐α‐elapitoxin‐1. Nonetheless, fibroblast growth factor signaling and heterotrimeric G‐protein signaling pathways were found to be uniquely expressed in Nn‐α‐elapitoxin‐1‐treated PC‐12 cells and not in mouse 2.5S‐nerve growth factor ‐treated cells. The TrkA binding region of Nn‐α‐elapitoxin‐1 may be developed as a peptide‐based drug prototype for the treatment of major central neurodegenerative diseases. image Read the Editorial Highlight for this article on page 599.
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... Surprisingly, Wnt4 gene expression was increased in the mutant at all three time points although the increase was only statistically significant on GD7.5 (Fig. 4, p < 0.0001). Similarly, the expression of amphiregulin (Areg) and brain-derived neurotrophic factor (Bdnf), both of which are important progesterone regulated genes for mammary cell growth 21,42,43 , were also significantly upregulated in the AF1_FFF mammary tissue in early gestation (Fig. 4). However, AF1_FFF mutations had no significant effect on the expression of Pgr or Esr1 although Pgr expression on GD7.5 showed a trend of upregulation (p > 0.05). ...
Activation function 1 of progesterone receptor is required for mammary development and regulation of RANKL during pregnancy
Article
Full-text available
  • Jul 2022
Progesterone receptor (PGR) is a member of the nuclear receptor superfamily of transcription factors. It is critical for mammary stem cells expansion, mammary ductal branching and alveologenesis. The transcriptional activity of PGR is mainly mediated by activation functions AF1 and AF2. Although the discovery of AF1 and AF2 propelled the understanding of the mechanism of gene regulation by nuclear receptors, their physiological roles are still poorly understood. This is largely due to the lack of suitable genetic models. The present study reports gain or loss of AF1 function mutant mouse models in the study of mammary development. The gain of function mutant AF1_QQQ exhibits hyperactivity while the loss of function mutant AF1_FFF shows hypoactivity on mammary development. However, the involvement of AF1 is context dependent. Whereas the AF1_FFF mutation causes significant impairment in mammary development during pregnancy or in response to estrogen and progesterone, it has no effect on mammary development in nulliparous mice. Furthermore, Rankl, but not Wnt4 and Areg is a major target gene of AF1. In conclusion, PGR AF1 is a pivotal ligand-dependent activation domain critical for mammary development during pregnancy and it exerts gene specific effect on PGR regulated genes.
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... Furthermore, TrkA is a therapeutic target in oncology and Trk inhibitors are being tested in several clinical trials [16][17][18][19]. In breast cancer, TrkA has been shown to stimulate tumour cell invasion and metastasis through the activation of ERK, PI3K, SRC and AKT signaling pathways [10,[20][21][22]. TrkA (NTRK1) gene overexpression has also been shown to occur preferentially in invasive tumours [23]. ...
The Receptor Tyrosine Kinase TrkA Is Increased and Targetable in HER2-Positive Breast Cancer
Article
Full-text available
  • Sep 2020
The tyrosine kinase receptor A (NTRK1/TrkA) is increasingly regarded as a therapeutic target in oncology. In breast cancer, TrkA contributes to metastasis but the clinicopathological significance remains unclear. In this study, TrkA expression was assessed via immunohistochemistry of 158 invasive ductal carcinomas (IDC), 158 invasive lobular carcinomas (ILC) and 50 ductal carcinomas in situ (DCIS). TrkA was expressed in cancer epithelial and myoepithelial cells, with higher levels of TrkA positively associated with IDC (39% of cases) (p < 0.0001). Interestingly, TrkA was significantly increased in tumours expressing the human epidermal growth factor receptor-2 (HER2), with expression in 49% of HER2-positive compared to 25% of HER2-negative tumours (p = 0.0027). A panel of breast cancer cells were used to confirm TrkA protein expression, demonstrating higher levels of TrkA (total and phosphorylated) in HER2-positive cell lines. Functional investigations using four different HER2-positive breast cancer cell lines indicated that the Trk tyrosine kinase inhibitor GNF-5837 reduced cell viability, through decreased phospho-TrkA (Tyr490) and downstream AKT (Ser473) activation, but did not display synergy with Herceptin. Overall, these data highlight a relationship between the tyrosine kinase receptors TrkA and HER2 and suggest the potential of TrkA as a novel or adjunct target for HER2-positive breast tumours.
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Nerve Growth Factor and the Role of Inflammation in Tumor Development
Article
Full-text available
  • Jan 2024
  • CURR ISSUES MOL BIOL
Nerve growth factor (NGF) plays a dual role both in inflammatory states and cancer, acting both as a pro-inflammatory and oncogenic factor and as an anti-inflammatory and pro-apoptotic mediator in a context-dependent way based on the signaling networks and its interaction with diverse cellular components within the microenvironment. This report aims to provide a summary and subsequent review of the literature on the role of NGF in regulating the inflammatory microenvironment and tumor cell growth, survival, and death. The role of NGF in inflammation and tumorigenesis as a component of the inflammatory system, its interaction with the various components of the respective microenvironments, its ability to cause epigenetic changes, and its role in the treatment of cancer have been highlighted in this paper.
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Electroacupuncture promotes apoptosis and inhibits axonogenesis by activating p75 neurotrophin receptor for triple-negative breast xenograft in mice
Article
  • Jun 2022
  • J CHEM NEUROANAT
Purpose The aim of this study was to investigate the anti-tumor effect of electroacupuncture (EA) on mice bearing breast tumors by regulating p75 neurotrophin receptor (p75NTR) and remodelling intratumoral innervation. Methods: Female BALB/c mice were implanted with 4T1 breast tumor cells to establish a murine mammary cancer model. Tumor volume and weight were measured to evaluate tumor growth. Cell apoptosis was assessed by TUNEL assay. The relative expression of p75NTR, TrkA, TrkB, NGF and proNGF were detected by immunohistochemistry. Neurotransmitter and neurotrophin were detected by enzyme-linked immunosorbent assay. Intratumoral innervation was confirmed by β3-tubulin and TH labeling immunohistochemistry. The antagonist TAT-Pep5 was employed to determine if the effects of EA on tumor growth and cell apoptosis were mediated by p75NTR. Results: Peritumoral EA alleviated tumor growth especially after 14 days of intervention. Apoptosis index in the tumor tissue was obviously decreased after EA. Meanwhile, EA intervention significantly upregulated the expression of p75NTR and proNGF, along with a decline in the tumor growth and an increase in the cell apoptosis. Besides, EA reduced local sympathetic innervation and downregulated sympathetic neurotransmitter NE level in the local tumor. Furthermore, p75NTR antagonist alleviated EA-mediated cell apoptosis and intratumoral innervation. Conclusions: One mechanism of EA intervention for alleviating tumor progression is mediated by p75NTR to promote apoptosis and decrease intratumoral axonogenesis in the tumor microenvironment.
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Role of insulin-like growth factors and the type I insulin-like growth factor receptor in the estrogen-stimulated proliferation of human breast cancer cells.
Article
Full-text available
  • Dec 1990
Estrogen sensitizes the MCF-7 estrogen-responsive breast cancer cell line to the mitogenic effect of insulin and the insulin-like growth factors (IGFs). This sensitization is specific for estrogen and occurs at physiological concentrations of estradiol. Dose-response experiments with insulin, IGF-I, and IGF-II suggested that the sensitization is mediated through the type I IGF receptor. Binding experiments with 125I-IGF-I and hybridization of a type I IGF receptor probe to RNA showed that the levels of the type I IGF receptor and its mRNA are increased 7- and 6.5-fold, respectively, by estradiol. IGF-I and estradiol had similar synergistic effects on other estrogen-responsive breast cancer cell lines, but IGF-I alone increased the proliferation of the MDA MB-231 cell line which is not responsive to estrogens. These experiments suggest that an important mechanism by which estrogens stimulate the proliferation of hormone-dependent breast cancer cells involves sensitization to the proliferative effects of IGFs and that this may involve regulation of the type I IGF receptor.
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Nerve Growth Factor Stimulates Proliferation and Survival of Human Breast Cancer Cells through Two Distinct Signaling Pathways
Article
Full-text available
  • May 2001
We show here that the neurotrophin nerve growth factor (NGF), which has been shown to be a mitogen for breast cancer cells, also stimulates cell survival through a distinct signaling pathway. Breast cancer cell lines (MCF-7, T47-D, BT-20, and MDA-MB-231) were found to express both types of NGF receptors: p140trkA and p75NTR. The two other tyrosine kinase receptors for neurotrophins, TrkB and TrkC, were not expressed. The mitogenic effect of NGF on breast cancer cells required the tyrosine kinase activity of p140trkA as well as the mitogen-activated protein kinase (MAPK) cascade, but was independent of p75NTR. In contrast, the anti-apoptotic effect of NGF (studied using the ceramide analogue C2) required p75NTR as well as the activation of the transcription factor NF-kB, but neither p140trkA nor MAPK was necessary. Other neurotrophins (BDNF, NT-3, NT-4/5) also induced cell survival, although not proliferation, emphasizing the importance of p75NTR in NGF-mediated survival. Both the pharmacological NF-κB inhibitor SN50, and cell transfection with IkBm, resulted in a diminution of NGF anti-apoptotic effect. These data show that two distinct signaling pathways are required for NGF activity and confirm the roles played by p75NTR and NF-κB in the activation of the survival pathway in breast cancer cells.
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Physiology of the Neurotrophins
Article
  • Jan 1996
The neurotrophins are a small group of dimeric proteins that profoundly affect the development of the nervous system of vertebrates. Recent studies have established clear correlations between the survival requirements for different neurotrophins of functionally distinct subsets of sensory neurons. The biological role of the neurotrophins is not limited to the prevention of programmed cell death of specific groups of neurons during development. Neurotrophin-3 in particular seems to act on neurons well before the period of target innervation and of normally occuning cell death. In animals lacking functional neurotrophin or receptor genes, neuronal numbers do not seem to be massively reduced in the CNS, unlike in the PNS. Finally, rapid actions of neurotrophins on synaptic efficacy, as well as the regulation of their mRNAs by electrical activity, suggest that neurotrophins might play important roles in regulating neuronal connectivity in the developing and in the adult central nervous system.
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Regression Models and Life Table
Article
  • Jan 1972
The analysis of censored failure times is considered. It is assumed that on each individual are available values of one or more explanatory variables. The hazard function (age-specific failure rate) is taken to be a function of the explanatory variables and unknown regression coefficients multiplied by an arbitrary and unknown function of time. A conditional likelihood is obtained, leading to inferences about the unknown regression coefficients. Some generalizations are outlined. LIFEtables are one of the oldest statistical techniques and are extensively used by medical statisticians and by actuaries. Yet relatively little has been written about their more formal statistical theory. Kaplan and Meier (1958) gave a comprehensive review of earlier work and many new results. Chiang in a series of papers has, in particular, explored the connection with birth-death processes; see, for example, Chiang (1968). The present paper is largely concerned with the extension of the results of Kaplan and Meier to the comparison of life tables and more generally to the incorporation of regression-like arguments into life-table analysis. The arguments are asymptotic but are relevant to situations where the sampling fluctuations are large enough to be of practical importance. In other words, the applications are more likely to be in industrial reliability studies and in medical statistics than in actuarial science. The procedures proposed are, especially for the two-sample problem, closely related to procedures for combining contingency tables; see Mantel and Haenzel (1959), Mantel (1963) and, especially for the application to life tables, Mantel (1966). There is also a strong connection with a paper read recently to the Society by R. and J. Peto (1972). We consider a population of individuals; for each individual we observe either the time to "failure" or the time to ccloss" or censoring. That is, for the censored individuals we know only that the time to failure is greater than the censoring time. Denote by T a random variable representing failure time; it may be discrete or continuous. Let F(t) be the survivor function, %(t) = pr (T2 t)
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A study in contrasts
Article
  • Jul 1968
  • PATTERNS PREJUDICE
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Regulation of Growth by a Nerve Growth Factor-like Protein Which Modulates Paracrine Interactions between a Neoplastic Epithelial Cell Line and Stromal Cells of the Human Prostate
Article
  • Jul 1991
Nerve growth factor-like substance(s) were identified in both conditioned media of a human prostatic tumor epithelial cell line (TSU-pr1) and a human prostatic stromal cell line (HPS) by Western blot analysis and bioassay of neurite outgrowth of PC12 cells. Nerve growth factor-beta (NGF) immunofluorescence was also localized to secretory vesicles in the cytoplasm of both the TSU-pr1 and HPS cells. Western blot of the TSU-pr1 and HPS cell-secreted protein identified an Mr 65,000 major protein which immunoreacted with murine NGF antibody. NGF Western blot of HPS cell-secreted protein also identified an Mr 42,000 minor band under reduced and nonreduced conditions and an Mr 61,000 minor band under reduced conditions. The secreted protein from the TSU-pr1 cells (50 micrograms/ml) and HPS (50 micrograms/ml), as well as murine NGF (50 ng/ml) or human recombinant NGF (50 ng/ml), stimulated neurite outgrowth from PC12 cells. This neurite outgrowth activity was partially inhibited by treatment with NGF antibody. Neither the serum containing growth medium nor bovine serum albumin (50 micrograms/ml) stimulated neurite outgrowth. The NGF-like secretory protein appeared to play a role in the paracrine regulation of prostatic growth between TSU-pr1 cells and HPS cells. The relative growth of TSU-pr1 cells, as indicated by [3H]thymidine incorporation, in response to HPS secretory protein was stimulated 2.8-fold in a dose-dependent manner. In the converse interaction, the relative growth of HPS cells in response to TSU-pr1 secretory protein was stimulated 1.8-fold in a dose-dependent manner. Immunoneutralization of TSU-pr1 and HPS secretory protein was performed with antibody against NGF, acidic fibroblast growth factor, and basic fibroblast growth factor. Removal of the NGF-like protein from the maximal stimulatory dose of TSU-pr1 secretory protein (100 micrograms/ml) with NGF antibody reduced HPS proliferation to 52% of maximal levels, and immunoneutralization of the NGF-like protein in the maximal stimulatory dose of HPS secretory protein (20 micrograms/ml) also reduced TSU-pr1 proliferation to 16% of maximal levels. Addition of normal rabbit serum or prior immunoprecipitation of either TSU-pr1 or HPS secretory protein with antibody against acidic fibroblast growth factor and basic fibroblast growth factor did not inhibit the proliferation of either cell type. These results suggest that TSU-pr1 tumor cells and HPS cells secrete NGF-like protein(s) which modulate their paracrine interactive growth in vitro.
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Prognostic significance of IGF1 receptors in human breast cancer
Article
  • Dec 1990
Overall survival (OS) and relapse free survival (RFS) were studied in 297 patients according to the presence of insulin-like growth factor 1 receptors (IGF1-R). All the patients were surgically treated for locoregional disease in the same institution from January 1986. The median duration of follow-up was 40 months. RFS was better in patients with IGF1-R in their tumors as assessed by actuarial survival (P = 0.014) as well as Cox analysis (P = 0.016). OS was better in IGF1-R positive tumors studied by actuarial (P = 0.007) as well as Cox analysis (P = 0.010). By Cox analysis the other prognostic factors on RFS were estrogen receptor (P = 0.002), progesterone receptor (P = 0.002), axillary node metastases (P = 0.032), histoprognostic grading (GHP) according to the standard of Scarff and Bloom (P = 0.004), and tumor diameter (P = 0.019). The other prognostic factors on OS (Cox analysis) were estrogen receptor (P = 0.001), axillary node metastases (P = 0.010), GHP (P = 0.009), progesterone receptor (P = 0.012), and tumor diameter (P = 0.007). When combining IGF1-R, GHP, and axillary node metastases, it appeared that IGF1-R, GHP, and axillary node metastases had independent prognostic significance. In this prospective study IGF1-R had a prognostic significance on RFS as well as on OS studied by actuarial as well as Cox analysis.
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Role of IGFs and IGF receptor in the estrogen stimulated proliferation of human breast cancer cells
Article
  • Jan 1991
Estrogen sensitizes the MCF-7 estrogen-responsive breast cancer cell line to the mitogenic effect of insulin and the insulin-like growth factors (IGFs). This sensitization is specific for estrogen and occurs at physiological concentrations of estradiol. Dose-response experiments with insulin, IGF-I, and IGF-II suggested that the sensitization is mediated through the type I IGF receptor. Binding experiments with 125I-IGF-I and hybridization of a type I IGF receptor probe to RNA showed that the levels of the type I IGF receptor and its mRNA are increased 7- and 6.5-fold, respectively, by estradiol. IGF-I and estradiol had similar synergistic effects on other estrogen-responsive breast cancer cell lines, but IGF-I alone increased the proliferation of the MDA MB-231 cell line which is not responsive to estrogens. These experiments suggest that an important mechanism by which estrogens stimulate the proliferation of hormone-dependent breast cancer cells involves sensitization to the proliferative effects of IGFs and that this may involve regulation of the type I IGF receptor.
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The importance of histologic grade in long-term prognosis of breast cancer: a study of 1,010 patients, uniformly treated at the Institut Gustave-Roussy
Article
  • Oct 1987
In a study of 1,010 patients with solitary, unilateral, nonmetastatic breast cancer, the histologic grade, assessed by a multifactorial analysis (Cox model) to study its significance with other prognostic factors, was found to be an important, independent factor. For 612 operable patients, two laboratory characteristics, the number of histologically positive nodes and the histologic grade, were the most valuable predictors. These two factors alone form a predictive index that may be an excellent and simple guide for the clinical decision of subsequent therapy. For 398 patients with inoperable breast cancer (ie, tumor greater than or equal to 7 cm, N2-3, inflammatory, skin fixation, and clinically rapidly growing forms), the histologic grade (performed on drill or cutting needle biopsy) was again a most important (and with inflammatory forms the most important) predictor of prognosis in these patients. Our data support that performing our modified histoprognostic grading of Scarff and Bloom is simple, reproducible, incurs no additional cost, may be carried out in the simplest histology laboratory, and finally, defines an important risk factor in all patients. It should be routine for all breast cancer specimens. Furthermore, studies of adjuvant therapy should stratify patients for this variable.
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Nerve Growth Factor in Mouse Milk during Early Lactation: Lack of Dependency on Submandibular Salivary Glands
Article
  • Oct 1985
Using a specific and sensitive nerve growth factor radioimmunoassay we show measurable quantities of beta nerve growth factor in mouse milk during the period of early lactation. Partial purification by cationic exchange resin yielded a preparation which exhibited biological activity in a PC-12 cell bioassay system. Submandibular-sublingual sialoadenectomy had no influence on the breast milk NGF concentrations. These results support the presence of bioactive NGF in mouse milk during early lactation, but do not clarify the source.
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