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NDGA inhibits PC3 xenografts growth and metastasis. (A) Immunohistochemistry staining of NRP1 in the tumor tissues of subcutaneous xenograft model. Scale bar: 100 ?m. (B) Representative bioluminescence images showing the metastatic sites in intravenous injection model. Mice of three groups (vehicle, 50 mg/kg NDGA, 100 mg/kg NDGA) were monitored after 14 days and 28 days drug treatments. (C) Quantification of bioluminescence signal within the region of interest at day 14 and day 28. Data show mean ? S.E (n = 5). *p < 0.05, **p < 0.01. (D) Quantification of bioluminescence signal of lungs at day 28. Data show mean ? S.E (n = 5). **p < 0.01. (E) Representative bioluminescence images of lungs with metastatic sites.
Source publication
Tumor metastasis is a major cause leading to the deaths of cancer patients. Nordihydroguaiaretic acid (NDGA) is a natural product that has been demonstrated to show therapeutic values in multiple diseases. In this study, we report that NDGA can inhibit cell migration and tumor metastasis via a novel mechanism. NDGA suppresses NRP1 function by downr...
Contexts in source publication
Context 1
... we inoculated tumor cells subcutaneously into the flank of nude mice to test whether NDGA administration affects NRP1 expression in vivo. Results show that NRP1 expression level in tumor tissues are lower in NDGA treated groups ( Figure 6A), consistent with our in vitro results. Besides, we also observed compromised tumor growth in NDGA treated groups (Supplementary Figure S2). ...
Context 2
... show that NDGA treatment significantly suppresses tumor metastasis in whole body range ( Figure 6B, 6C). Further, after 4 weeks the mice were sacrificed and metastatic nodules on lungs were observed by bioluminescence. ...
Context 3
... after 4 weeks the mice were sacrificed and metastatic nodules on lungs were observed by bioluminescence. Again, we found less metastatic sites in NDGA treated mice than control ( Figure 6D, 6E). By these results, we can further confirm the inhibitory effect of NDGA on tumor metastasis. ...
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A bstract
Cell migration is involved in key phenomena in biology, ranging from development to cancer. Fibroblasts move between organs in 3D polymeric networks. So far, motile cells were mainly tracked in vitro on Petri dishes or on coverslips, i.e. 2D flat surfaces, which made the extrapolation to 3D physiological environments difficult. We therefo...
Citations
... A recent study found that miR-124-3p regulates NRP-1 expression in MDA-MB-231 cells, and reducing the expression of NRP-1 in mouse 4T1 cells reduced liver metastasis via the TGFβ pathway [32]. In prostate cancer PC3 cells, NRP-1 downregulation was correlated with the impaired ability of the cells to metastasize to the lungs upon nordihydroguaiaretic acid treatment [33]. ...
Breast cancer (BC) metastasis remains a leading cause of female mortality. Neuropilin-1 (NRP-1) is a glycoprotein receptor that plays ligand-dependent roles in BC. Clinical studies indicate its correlation with metastatic disease; however, its functional role in BC metastasis remains uncertain. CRISPR-Cas9 was used to knockout the NRP-1 gene in MDA-MB-231 BC cells, and the effects on metastasis were determined using an orthotopic mouse engraftment model. NRP-1 expression in knockout cells was rescued using a recombinant cDNA with a silent mutation in the sgRNA target-adjacent PAM sequence. Differentially expressed genes between NRP-1 knockout and control cells were determined using whole-transcriptome sequencing and validated using real-time PCR. NRP-1KO cells showed a pronounced reduction in the metastasis to the lungs. KEGG pathway analysis of the transcriptome data revealed that PI3K and ECM receptor interactions were among the top altered pathways in the NRP-1KO cells. In addition, reduction in metastasis enhancers proteins, Integrin-β3 and Tenascin-C, and genes CCL20 and FN1 and upregulation of metastasis suppressor genes, ACVRL and GPX3 in NRP-1KO were detected. These findings provide evidence for a functional role for NRP-1 in BC metastasis, supporting further exploration of NRP-1 and the identified genes as targets in treating metastatic BC.
... NDGA was suggested to suppress, via its antioxidant action, the pathogenesis of cardiovascular or other diseases that ROS contribute to and aggravate [21]. In addition, this compound may suppress several types of tumor cell growth [22][23][24][25][26]. Although little is known about the anticancer action of NDGA, its inhibitory activity of several enzymes, such as lipoxygenase, may contribute to this. ...
Background
Methylglyoxal (MGO) is a known toxic byproduct of glycolysis, with MGO-induced cytotoxicity believed to contribute to the pathogenesis of several diseases. Glyoxalase I (GLO1) is a key enzyme for eliminating MGO in mammalian cells, therefore, compounds affecting GLO1 activity are potential therapeutic agents for MGO-induced disorders. Previously, we found nordihydroguaiaretic acid (NDGA) as a potent GLO1 inhibitor.
Methods
The inhibitory characteristics of NDGA were determined spectrophotometrically with recombinant GLO1. NDGA-induced growth-inhibition and accumulation of MGO-derived advanced glycation end products (AGEs) were examined in EA.hy926 cells.
Results
NDGA showed significant inhibition of GLO1 enzymatic activity in a dose-dependent manner. Its Ki value was estimated to be 146-fold lower than that of myricetin, a known GLO1 inhibitor. The co-addition of MGO with NDGA to the cells resulted in significant growth inhibition, suggesting that MGO accumulation, sufficient to affect cell growth, was caused by NDGA inhibiting GLO1. These findings were supported by the observations that the addition of aminoguanidine, a typical MGO scavenger, significantly reversed cell-growth inhibition by co-addition of MGO with NDGA, and that an increase in intracellular MGO-derived AGEs was observed during incubation with the co-addition of MGO with NDGA.
Conclusion
NDGA was found to be a novel and potent inhibitor of GLO1. The co-addition of NDGA with MGO to the cells resulted in increased intracellular MGO accumulation followed by enhanced cell-growth inhibition.
... Furthermore, co-targeting NRP1 and NRP2 expression effectively inhibited secondary site metastasis compared to control animals. Given both NRP1 and NRP2 are known to modulate primary and secondary tumour microenvironments by interacting with integrins to remodel the tumoral ECM [20], [45], [47], of which FN is known as a major component [48], it follows that the impaired tumour growth exhibited following NRP codepletion likely arises as a result of perturbations in EDA-FN fibril assembly and deposition. Indeed, EDA-FN has been demonstrated to facilitate tumour growth and invasiveness by promoting matrix stiffness, sustaining tumour-induced angiogenesis and lymphangiogenesis via VEGF-A165 [49] and VEGF-C respectively [50]. ...
Neuropilin (NRP) expression is highly correlated with poor outcome in multiple cancer subtypes. As known co-receptors for vascular endothelial growth factor receptors (VEGFRs), core drivers of angiogenesis, past investigations have alluded to their functional roles in facilitating tumorigenesis by promoting invasive vessel growth. Despite this, it remains unclear as to whether NRP1 and NRP2 act in a synergistic manner to enhance pathological angiogenesis. Here we demonstrate, using NRP1ECKO, NRP2ECKO and NRP1/NRP2ECKO mouse models, that maximum inhibition of primary tumour development and angiogenesis is only achieved when both endothelial NRP1 and NRP2 are targeted simultaneously. Metastasis and secondary site angiogenesis were also significantly inhibited in NRP1/NRP2ECKO animals. Mechanistic studies revealed that co-depleting NRP1 and NRP2 in mouse-microvascular endothelial cells (ECs) stimulates rapid shuttling of VEGFR-2 to Rab7+ endosomes for proteosomal degradation. Our results highlight the importance of targeting both NRP1 and NRP2 to modulate tumour angiogenesis.
... In addition, NDGA inhibits growth and induces cell cycle arrest at the G1 phase in cervical cancer-derived cell line SiHa by regulating cell cycle kinase inhibitor p21 and promoting the acetylation of histone H3 [27]. In the prostate cancer PC3 cell line, NDGA impairs cell migration and tumor metastasis in nude mice via suppressing the function of single-pass transmembrane protein neuropilin 1 [28]. Regarding the toxic effect of NDGA in no cancer cells, it has been reported that only high concentrations of NDGA (100 µM) induce lipid peroxidation, DNA double-stranded breaks, and cell death in rat hepatocytes [29] and do not affect the viability of normal cells and peripheral blood mononuclear cells (PBMC) at 3, 10, 30, and 60 µM [30]. ...
Medulloblastoma is a common malignant brain tumor in the pediatric age. The current therapeutics present serious collateral effects. Polyphenols α-mangostin and nordihydroguaiaretic acid (NDGA) exert potent antitumoral activity in different cancer models, although their antitumoral effects have not been described in medulloblastoma cells yet. This study aimed to examine the proapoptotic effects of these polyphenols on human medulloblastoma cells. Medulloblastoma cell line Daoy was incubated with increasing concentrations of α-mangostin or NDGA for 24 h. The cell viability was analyzed using crystal violet and trypan blue dyes. Determination of the glutathione (GSH)/glutathione disulfide (GSSG) ratio and levels of carbonylated proteins was performed to evaluate the oxidative stress. Cell cycle progression and induction of cell death by fluorochrome-couple and TUNEL assays were evaluated using flow cytometry assays. Individual treatments with α-mangostin or NDGA decreased the viability of Daoy cells in a dose-dependent manner, inducing G2/M and S-G2/M cell cycle arrest, respectively. Both polyphenols induced cell death and increased oxidative stress. Very interestingly, α-mangostin showed more potent effects than NDGA. Our results indicate that α-mangostin and NDGA exert important cytostatic and cytotoxic effects in the Daoy cell line. These data highlight the potential usefulness of these compounds as an alternative strategy in medulloblastoma treatment.
... We were able to identify genes coding for cell surface-bound proteins, which can potentially be explored as targets for radiolabeled monoclonal antibodies for positron emission tomography (PET)-based detection of metastatic prostate cancer. These markers include ADAM15 [48], CD276 [49], NRP1 [52,53], SCARB1 [54], and PLXNA3 [56], all of which have been reported to be overexpressed in metastatic PrCa. Elevated expression of genes such as ABCC5 [50], LRFN1 [59], ELOVL6 [58], and HTR2B [61] have been associated with metastasis in other cancer types. ...
This manuscript demonstrates how integrated bioinformatic and statistical reanalysis of publicly available genomic datasets can be utilized to identify molecular pathways and biomarkers that may be clinically relevant to metastatic prostate cancer (mPrCa) progression. The most notable observation is that the transition from primary prostate cancer to mPrCa is characterized by upregulation of processes associated with DNA replication, metastasis, and events regulated by the serine/threonine kinase PLK1. Moreover, our analysis also identified over-expressed genes that may be exploited for potential targeted therapeutics and minimally invasive diagnostics and monitoring of mPrCa. The primary data analyzed were two transcriptional datasets for tissues derived from normal prostate, primary prostate cancer, and mPrCa. Also incorporated in the analysis were the transcriptional, gene dependency, and drug response data for hundreds of cell lines, including those derived from prostate cancer tissues.
... NRPs are highly conserved, multifunctional transmembrane proteins that are unique to vertebrates and are involved in various physiological and pathological processes in the body [34,35]. In mammals, there are two isoforms of NRPs (NRP1 and NRP2) that are functionally distinct and complementary. ...
Background:
Neurovascular-related genes have been implicated in the development of cancer. Studies have shown that a high expression of neuropilins (NRPs) promotes tumourigenesis and tumour malignancy.
Method:
A multidimensional bioinformatics analysis was performed to examine the relationship between NRP genes and prognostic and pathological features, tumour mutational burden (TMB), microsatellite instability (MSI), and immunological features based on public databases and find the potential prognostic value of NRPs in pancancer.
Results:
Survival analysis revealed that a low NRP1 expression in adrenocortical carcinoma (ACC), cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC), low-grade glioma (LGG), and stomach adenocarcinoma (STAD) was associated with poor prognosis. A high NRP2 expression in bladder urothelial carcinoma (BLCA), kidney renal papillary cell carcinoma (KIRP), and mesothelioma (MESO) was associated with poor prognosis. Moreover, NRP1 and NRP2 were associated with TMB and MSI. Subsequent analyses showed that NRP1 and NRP2 were correlated with immune infiltration and immune checkpoints. Genome-wide association analysis revealed that the NRP1 expression was strongly associated with kidney renal clear cell carcinoma (KIRC), whereas the NRP2 expression was closely associated with BLCA. Ultimately, NRP2 was found to be involved in the development of BLCA.
Conclusions:
Neurovascular-related NRP family genes are significantly correlated with cancer prognosis, TME, and immune infiltration, particularly in BLCA.
... Baicalein, NDGA and zileuton, the drugs of natural origin used in this work, possess the capacity to inhibit lipoxygenase pathways, which results in their antitumor activity. NDGA, a non-selective inhibitor of the LOX pathway, contributes to increased apoptosis in different cell lines including neuroblastoma [22], gastrointestinal cancer [2], as well as suppresses migration and metastasis of prostate cancer cells [23]. However, in some cases, the antitumor effect of NDGA was probably not associated with inhibition of lipoxygenases [22]. ...
Several biochemical mechanisms, including the arachidonic acid cascade and activation of nicotinic acetylcholine receptors (nAChRs), are involved in increased tumor survival. Combined application of inhibitors acting on these two pathways may result in a more pronounced antitumor effect. Here, we show that baicalein (selective 12-lipoxygenase inhibitor), nordihydroguaiaretic acid (non-selective lipoxygenase inhibitor), and indomethacin (non-selective cyclooxygenase inhibitor) are cytotoxic to Ehrlich carcinoma cells in vitro. Marine snail α-conotoxins PnIA, RgIA and ArIB11L16D, blockers of α3β2/α6β2, α9α10 and α7 nAChR subtypes, respectively, as well as α-cobratoxin, a blocker of α7 and muscle subtype nAChRs, exhibit low cytotoxicity, but enhance the antitumor effect of baicalein 1.4-fold after 24 h and that of nordihydroguaiaretic acid 1.8–3.9-fold after 48 h of cell cultivation. α-Conotoxin MII, a blocker of α6-containing and α3β2 nAChR subtypes, increases the cytotoxic effect of indomethacin 1.9-fold after 48 h of cultivation. In vivo, baicalein, α-conotoxins MII and PnIA inhibit Ehrlich carcinoma growth and increase mouse survival; these effects are greatly enhanced by the combined application of α-conotoxin MII with indomethacin or conotoxin PnIA with baicalein. Thus, we show, for the first time, antitumor synergism of α-conotoxins and arachidonic acid cascade inhibitors.
... The tumor suppressor effect of miR-19b-3p could be reversed by overexpression of NRP1. NRP1 was overexpressed in various cancers including lung cancer, hepatocellular carcinoma, prostate cancer, and glioblastoma 27,28,29,30 . ...
Background Gastric cancer (GC) remains one of the most common digestive malignancies worldwide and ranked third causes of cancer-related death. Mounting evidence has revealed that miRNAs exert critical regulatory roles in GC development.
Methods Immunohistochemistry (IHC) and western blot assay were performed to determine the protein expression levels of neuropilin-1 (NRP1) and mRNA levels were confirmed by quantitative RT-PCR (qRT-PCR) in GC tissues. Kaplan–Meier analysis was performed to evaluate the prognostic value of NRP1 in GC. Knockdown of NRP1 was conducted to analyse its function in vitro and vivo. Luciferase reporter assay, western blot and qRT-qPCR were employed to identify the miRNAs which directly targeted NRP1. Furthermore, Bioinformatics analysis and experimental verification were used to explore the potential molecular mechanism and signaling pathway.
Results In the current study, we revealed that neuropilin-1 (NRP1) was highly expressed in GC tumor tissues and was associated with poor prognosis in GC patients. NRP1 knockdown inhibited GC cell growth, migration and invasion in vitro, while suppressed GC xenograft tumor development in vivo. Bioinformatics analysis predicted that miR-19b-3p down-regulated NRP1 expression by targeting its 3’-UTR. Functional assay demonstrated that miR-19b-3p inhibited GC cell growth, migration and invasion via negatively regulating NRP1. Overexpression NRP1 partially reversed the regulatory effect of miR-19b-3p. Moreover, we showed that miR-19b-3p/NRP1 axis regulated the epithelial-to-mesenchymal transition and focal adhesion in GC, which might contribute the GC development and progression.
Conclusions Taken together, our findings suggest a regulatory network of miR-19b-3p/NRP1 in GC development. The miR-19b-3p/NRP1 axis might be further explored as a potential diagnostic and therapeutic target in GC.
... It shows antiangiogenic effects by downregulation of the JAK-signal transducer and activator of transcription 3 (STAT3) pathway and STAT3-regulated genes, such as VEGF, Bcl-xL, and cyclin D1 [68]. Nordihydroguaiaretic acid, a phenolic compound extracted from creosote bush (Larrea tridentate), inhibited the NRP1 expression in PC3 cells, which is a co-receptor of VEGF [69]. Terrestrosin D is a major steroidal saponin derived from Tribulus terrestris L., which is a medicinal plant distributed widely in the Mediterranean. ...
Prostate cancer is the third most common cancer worldwide, and the burden of the disease is increased. Although several chemotherapies have been used, concerns about the side effects have been raised, and development of alternative therapy is inevitable. The purpose of this study is to prove the efficacy of dietary substances as a source of anti-tumor drugs by identifying their carcinostatic activities in specific pathological mechanisms. According to numerous studies, dietary substances were effective through following five mechanisms; apoptosis, anti-angiogenesis, anti-metastasis, microRNA (miRNA) regulation, and anti-multi-drug-resistance (MDR). About seventy dietary substances showed the anti-prostate cancer activities. Most of the substances induced the apoptosis, especially acting on the mechanism of caspase and poly adenosine diphosphate ribose polymerase (PARP) cleavage. These findings support that dietary compounds have potential to be used as anticancer agents as both food supplements and direct clinical drugs.
... Nordihydroguaiaretic acid can induce apoptosis and inhibit the growth of human lung cancer cells [191]. In addition, nordihydroguaiaretic acid can also inhibit cell migration and tumor metastasis in prostate cancer [192]. Proadifen, an inhibitor of cytochrome P450 monooxygenases, has anti-proliferative properties through influencing metabolic activity, cell number, and cell cycle progression in colon cancer [193]. ...
Non-small-cell lung cancer (NSCLC) is the predominant type of lung cancer in the world. Lung adenocarcinoma (LADC) and lung squamous cell carcinoma (LSCC) are subtypes of NSCLC. We usually regard them as different disease due to their unique molecular characteristics, distinct cells of origin and dissimilar clinical response. However, the differences of genetic and epigenetic progression mechanism between LADC and LSCC are complicated to analyze. Therefore, we applied systems biology approaches and big databases mining to construct genetic and epigenetic networks (GENs) with next-generation sequencing data of LADC and LSCC. In order to obtain the real GENs, system identification and system order detection are conducted on gene regulatory networks (GRNs) and protein-protein interaction networks (PPINs) for each stage of LADC and LSCC. The core GENs were extracted via principal network projection (PNP). Based on the ranking of projection values, we got the core pathways in respect of KEGG pathway. Compared with the core pathways, we found significant differences between microenvironments, dysregulations of miRNAs, epigenetic modifications on certain signaling transduction proteins and target genes in each stage of LADC and LSCC. Finally, we proposed six genetic and epigenetic multiple-molecule drugs to target essential biomarkers in each progression stage of LADC and LSCC, respectively.
