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A. CFSE cell proliferation assay showed that the rate of proliferation of untreated U87MG and U87-MCSF cells remained unaltered. B. Cell cycle analysis of U87MG and U87-MCSF cells after treatment with 5-FU for 24 h. Majority of cells were accumulated in G0/G1 phase in treated population of U87-MCSF cells. Statistical significance is denoted by * (p<0.05), ** (p<0.01) and *** (p<0.001).
Source publication
Macrophage colony stimulating factor (MCSF) regulates growth, proliferation and differentiation of haematopoietic cell lineages. Many cancers are known to secrete high level of MCSF, which recruit macrophages into the tumour micro-environment, supporting tumour growth. Herein, we report the cloning of MCSF and subsequent generation of U87MG express...
Citations
... The ATP-binding cassette subfamily B member 1 (ABCB1) gene, also known as multidrug resistance 1 (MDR1), codes for P-glycoprotein (P-gp), a carrier protein that transports a great variety of substrates and plays a crucial role in maintaining intracellular levels of numerous antineoplastic agents [13]. Although capecitabine has not been clearly identified as a substrate of P-gp, its expression has been related to resistance to 5-FU in modified cell lines, so its potential influence on the outcomes of therapy cannot be ruled out [14]. sion of 5,10-methylenetetrahydrofolate (5,10-MTHF) to dihydrofolate, a key component of the folate cycle. ...
Simple Summary
Colorectal cancer is one of the most prevalent neoplasms worldwide. Capecitabine is an oral fluoropyrimidine widely used to treat colorectal cancer in early and advanced stages. However, it shows high interindividual variability in its effectiveness and safety. This variability may be due to genetic variants in proteins involved in the pharmacokinetics and pharmacodynamics of the drug. Currently, only four variants of the DPYD gene are clinically relevant for the prediction of severe toxicity, and there are no validated predictive biomarkers of capecitabine effectiveness. Therefore, the search of potential predictive genetic biomarkers to personalize and optimize capecitabine therapy remains necessary. The aim of this study was to systematically review the literature published in the last 10 years on the influence of single-nucleotide polymorphisms in the main genes involved in capecitabine pharmacokinetics and pharmacodynamics on therapy outcomes in patients with colorectal cancer.
Abstract
The aim of this systematic review was to provide a comprehensive overview of the literature published in the last decade on the association of single-nucleotide polymorphisms in genes involved in the pharmacodynamic and pharmacokinetic pathways of capecitabine with treatment outcomes among colorectal cancer patients. A systematic search of the literature published in the last 10 years was carried out in two databases (Medline and Scopus) using keywords related to the objective. Quality assessment of the studies included was performed using an assessment tool derived from the Strengthening the Reporting of Genetic Association (STREGA) statement. Thirteen studies were included in this systematic review. Genes involved in bioactivation, metabolism, transport, mechanism of action of capecitabine, DNA repair, and folate cycle were associated with toxicity. Meanwhile, genes related to DNA repair were associated with therapy effectiveness. This systematic review reveals that several SNPs other than the four DPYD variants that are screened in clinical practice could have an impact on treatment outcomes. These findings suggest the identification of future predictive biomarkers of effectiveness and toxicity in colorectal cancer patients treated with capecitabine. However, the evidence is sparse and requires further validation.
... Aurora kinase A (AURKA), as an ATM kinase, was reported to cause radio-resistance through self-activation (De Bacco et al., 2016). Xenopus kinesin-like protein 2 (TPX2) was reported to promote tumor invasion and proliferation (Chockalingam and Ghosh, 2013). Notably, through protein interaction and co-expression networks, we speculate that AURKA, CCNA2, and TPX2 are ideal drug targets in GBM, which provide potential evidence for stem cell treatment in GBM. ...
The progression of most human cancers mainly involves the gradual accumulation of the loss of differentiated phenotypes and the sequential acquisition of progenitor and stem cell-like features. Glioblastoma multiforme (GBM) stem cells (GSCs), characterized by self-renewal and therapeutic resistance, play vital roles in GBM. However, a comprehensive understanding of GBM stemness remains elusive. Two stemness indices, mRNAsi and EREG-mRNAsi, were employed to comprehensively analyze GBM stemness. We observed that mRNAsi was significantly related to multi-omics parameters (such as mutant status, sample type, transcriptomics, and molecular subtype). Moreover, potential mechanisms and candidate compounds targeting the GBM stemness signature were illuminated. By combining weighted gene co-expression network analysis with differential analysis, we obtained 18 stemness-related genes, 10 of which were significantly related to survival. Moreover, we obtained a prediction model from both two independent cancer databases that was not only an independent clinical outcome predictor but could also accurately predict the clinical parameters of GBM. Survival analysis and experimental data confirmed that the five hub genes (CHI3L2, FSTL3, RPA3, RRM2, and YTHDF2) could be used as markers for poor prognosis of GBM. Mechanistically, the effect of inhibiting the proliferation of GSCs was attributed to the reduction of the ratio of CD133 and the suppression of the invasiveness of GSCs. The results based on an in vivo xenograft model are consistent with the finding that knockdown of the hub gene inhibits the growth of GSCs in vitro. Our approach could be applied to facilitate the development of objective diagnostic and targeted treatment tools to quantify cancer stemness in clinical tumors, and perhaps lead considerable benefits that could predict tumor prognosis, identify new stemness-related targets and targeted therapies, or improve targeted therapy sensitivity. The five genes identified in this study are expected to be the targets of GBM stem cell therapy.
... Activation of M-CSF pathways in cancer cells is correlated with the number of tumor-associated M-CSF receptor-positive M2 macrophages in patients with lung cancer [39]. M-CSF expressed by glioblastoma cells plays a role in resistance to the anticancer drug 5-FU [40]. M-CSF derived from glioblastoma also promotes angiogenesis by microglial release of insulin-like growth factor-binding protein 1 [41]. ...
Background:
Expression of Oct4 maintains cancer stem cell (CSC)-like properties in lung cancer cells and is correlated with poor prognosis of lung adenocarcinoma. M2-type tumor-associated macrophages (TAMs) promote cancer cell migration and metastasis. Tumor microenvironments promote monocyte differentiation into M2 TAMs via a complex cytokine-based connection. We explored the role of Oct4 in cytokine secretion in lung cancer and its impact on M2 TAM polarization.
Methods:
Monocytes co-cultured with the conditioned medium from Oct4-overexpressing lung cancer cells were used to investigate M2 TAM differentiation. The inflammatory factors in the conditioned medium of Oct4-overexpressing A549 cells were examined using human inflammation antibody arrays. The correlations of Oct4, macrophage colony-stimulating factor (M-CSF), and M2 TAMs were validated in lung cancer cells, syngeneic mouse lung tumor models, and clinical samples of non-small cell lung cancer (NSCLC).
Results:
Oct4-overexpressing A549 cells expressed elevated levels of M-CSF, which contributed to increased M2 macrophages and enhanced tumor migration. Overexpression of Oct4 enhanced tumor growth and reduced the survival of lung tumor-bearing mice, which was correlated with increased number of M2 macrophages in lung cancer. Notably, NSCLC patients with high expression levels of Oct4, M-CSF, and M2 TAMs had the poorest recurrence-free survival. A positive correlation between Oct4, M-CSF, and M2 TAMs was observed in the tumor tissue of NSCLC patient. Treatment with all-trans retinoic acid exerted anti-tumor effects and reduced M2 TAMs in tumor-bearing mice.
Conclusions:
Our results indicate that Oct4 expressed by lung cancer cells promotes M2 macrophage polarization through upregulation of M-CSF secretion, leading to cancer growth and metastasis. Our findings also implicate that the Oct4/M-CSF axis in M2 macrophage polarization may be potential therapeutic targets for lung cancer.
... Further, cytoskeleton staining of untreated and treated samples of U87MG and U87-MCSF cells was done using anti β-actin antibody (Figure 3b). The results obtained reinforce the morphological changes observed in MB staining [69]. Problems that arise in practice must be taken into account, where interference may occur which would transform the benefits of using these compounds into undesirable effects. ...
Methylene blue and proflavine are fluorescent dyes used to stain nucleic acid from the molecular level to the tissue level. Already clinically used for sentinel node mapping, detection of neuroendocrine tumors, methemoglobinemia, septic shock, ifosfamide-induced encephalopathy, and photodynamic inactivation of RNA viruses, the antimicrobial, anti-inflammatory, and antioxidant effect of methylene blue has been demonstrated in different in vitro and in vivo studies. Proflavine was used as a disinfectant and bacteriostatic agent against many gram-positive bacteria, as well as a urinary antiseptic involved in highlighting cell nuclei. At the tissue level, the anti-inflammatory effects of methylene blue protect against pulmonary, renal, cardiac, pancreatic, ischemic-reperfusion lesions, and fevers. First used for their antiseptic and antiviral activity, respectively, methylene blue and proflavine turned out to be excellent dyes for diagnostic and treatment purposes. In vitro and in vivo studies demonstrated that both dyes are efficient as perfusion and tissue tracers and permitted to evaluate the minimal efficient concentration in different species, as well as their pharmacokinetics and toxicity. This review aims to identify the optimal concentrations of methylene blue and proflavine that can be used for in vivo experiments to highlight the vascularization of the skin in the case of a perforasome (both as a tissue tracer and in vascular mapping), as well as their effects on tissues. This review is intended to be a comparative and critical presentation of the possible applications of methylene blue (MB) and proflavine (PRO) in the surgical field, and the relevant biomedical findings from specialized literature to date are discussed as well.
... 15 Additionally, recent reports have established that the upregulation of key ABC transporter genes (ABCG1 and ABCB1) imparts resistance to 5-FU in U87-MCSF cells. 16 Our data showed that the expression level of ABCG1 in cells irradiated by 12 C ions increased as the dose increased. Since the increased expression of various ABC transporters may lead to chemo-resistance, the expression levels of ABC transporters should be detected before treatment with both chemotherapy and heavy ion radiotherapy. ...
Objective
Heavy ions have contributed to tumor site-specific radiotherapy and are a major health risk for astronauts. The purpose of this study was to investigate the changes in gene expression in peripheral lymphocytes of cancer patients and astronauts exposed to ¹²C ions, and identify suitable molecular biomarkers for health monitoring. We also aimed to observe the effects of treatment and the level of damage, by comparing the transcriptional profiles of human lymphocyte cell lines exposed to ¹²C ion beams at doses of 0–2.0 Gy.
Materials and methods
A human lymphocyte cell line was irradiated with ¹²C ion beams at 0, 0.1, 0.5, and 2.0 Gy and transcriptional profiles were evaluated using the Agilent human gene expression microarray at 24 hours after irradiation. Differentially expressed genes were identified using a fold change of ≥2.0. Representative genes were further validated by RT-PCR. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses were performed to determine the roles of differentially expressed mRNAs.
Results
Based on the microarray assays, 1,113 genes were upregulated and 853 genes were downregulated in human lymphocyte cells irradiated with 0.1 Gy ¹²C ion beams compared with the control group, 1,095 genes were upregulated and 1,220 genes were downregulated in cells irradiated with 0.5 Gy ¹²C ion beams, and 1,055 genes were upregulated and 1,356 genes were downregulated in cells irradiated with 2.0 Gy. A total of 504 genes were differentially expressed in all irradiated groups, of which 88 genes were upregulated and 416 genes downregulated. Most of these altered genes were related to the cell cycle, apoptosis, signal transduction, DNA transcription, repair, and replication. The expression differences were further confirmed by RT-PCR for a subset of differentially expressed genes.
Conclusion
Differentially expressed genes between treatment and control groups at 24 hours post-irradiation increased as the radiation dose increased; upregulated genes gradually decreased and downregulated genes increased. Our data indicated that ¹²C ion beams could repress a number of genes in a dose-dependent manner, which might lead to the failure of multiple cellular biological functions.
... Aharinejad et al found that the high expression of cytoplasmic M-CSF in MDA-MB-231 breast cancer cells contributes to the invasion and metastasis of tumours in a mouse model (2). Similarly, M-CSF was revealed to play an important role in the resistance of 5-FU in U87MG glioblastoma (5). In addition, an M-CSF antibody was revealed to reverse the chemoresistance of MCF-7 cells (6). ...
... Murine double minute 2 (MDM2), is an oncogene that is upstream of HIF-1α and regulates the expression of HIF-1α (12,13). M-CSF was revealed to directly decrease the expression of MDM2, further contributing to drug resistance in tumour treatments (5). M-CSF may modulate the expression of HIF-1α, however, the mechanism is still unclear. ...
Macrophage colony-stimulating factor (M-CSF), a tumour marker, is related to tumour cell anti-apoptosis and drug resistance. However, the role of M-CSF in MCF-7 cells is unknown. In the present study, the effect and mechanism of M-CSF on hypoxia-inducible factor-1α (HIF-1α)/BCL2/adeno-virus E1B 19 kDa-interacting protein 3 (BNIP3)/Apoptosis Regulator BAX signalling in human breast cancer MCF-7 cells were investigated. Western blotting revealed that the expression of HIF-1α, BNIP3, Bax, caspase-3 and caspase-9 was lower in MCF-7-M cells compared to MCF-7 and MCF-7-C cells treated with adriamycin (ADM). Immunoprecipitation combined with western blotting was used to detect the interaction between Bcl-2 and BNIP3 or Bax protein. MCF-7-M cells had a higher amount of Bax binding to Bcl-2 compared to MCF-7 cells or MCF-7-C cells, while the amount of BNIP3 binding to Bcl-2 was decreased in MCF-7-M cells. Hoechst 33342 staining and flow cytometry were utilized to evaluate the effect of M‑CSF on apoptosis in MCF-7 cells treated with ADM. Compared to ADM-treated MCF-7 cells, the apoptotic rate of MCF-7-M cells was significantly decreased. These effects were dependent on the concentration of ADM. In conclusion, cytoplasmic M-CSF suppressed apoptosis by inhibiting the HIF-1α/BNIP3/Bax signalling pathway, which potentiated the dissociation of Bcl-2 from Bcl-2-BNIP3 compounds and the formation of Bcl-2-Bax compounds.
... Additionally, it is known that CSCs can effectively increase drug resistance through upregulating the expression of drug efflux transporter genes and various multidrug resistance genes (41). CSCs isolated from the sphere-forming culture of cancer cells were previously demonstrated to possess high expression of the ABCB1 gene and were identified to be significantly resistant to various chemotherapeutic agents (42). In the present study, the expression levels of the resistance-associated proteins MDR1, ABCG1, ABCG2 and RAD51 were significantly downregulated following inhibition of Hes1. ...
It has been considered that the neurogenic locus notch homolog protein (Notch) signaling pathway serves an essential role in cellular differentiation, proliferation and apoptosis. However, the function of the Notch signaling pathway in gastric cancer stem cells (GCSCs) and epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) sensitivity remains unclear. The present study aimed to delineate the role of the Notch1 signaling pathway in GCSCs and lapatinib sensitivity. Sphere-forming cells were separated from human gastric cancer MKN45 parental cells. The sphere-forming cells exhibited characteristics of CSCs and higher Notch1 expression compared with that of parental cells. To investigate the role of the Notch1 signaling pathway in GCSCs, the expression of transcription factor Hes1 (Hes1) was knocked down using small interfering RNA against Hes1. It was observed that Hes1 expression was significantly downregulated in knocked down cells. The inhibition of Hes1 suppressed the properties of CSCs, as indicated by significant decreases in the expression of the transcription factor sex determining region Y-box 2, epithelial cell adhesion molecule and the homeobox protein Nanog and reduced spheroid colony formation. In addition, epithelial-mesenchymal transition was significantly impaired in sphere-forming cells following Hes1 knockdown. Furthermore, the inhibition of Hes1 effectively enhanced lapatinib sensitivity in sphere-forming cells. These results suggest that sphere-forming gastric cancer cells possess the characteristics of CSCs, and that the Notch1 signaling pathway serves an essential role in the maintenance of CSCs and lapatinib sensitivity.
... It is extensively expressed in the gastrointestinal epithelium and might control the uptake of oral agents such as UFT from the gut [27]. Although fluoropyrimidines are not a known substrate of ABCB1, ABCB1 expression is induced along with 5-FU resistance in some cell lines [28], which suggests polymorphisms concerning ABCB1 activity might be related to efficacy of fluoropyrimidines. Otherwise, ABCB1 haplotype*1 was reportedly associated with capecitabine toxicity compared to variant haplotypes [29]. ...
Background
This study aimed to evaluate the efficacy of a high dose of oral tegafur-uracil (400 mg/m2) plus leucovorin with preoperative chemoradiation of locally advanced rectal cancer and to explore the impact of polymorphisms of cytochrome P 2A6 (CYP2A6), uridine monophosphate synthetase (UMPS), and ATP-binding cassette B1 (ABCB1) on clinical outcome. Methods
Patients with cT3 or cT4 rectal cancer were enrolled and were given tegafur-uracil 400 mg/m2/day and leucovorin 90 mg/m2/day for 7 days a week during preoperative chemoradiation (50.4 Gy/28 fractions) in this phase II trial. Primary endpoint was pathologic complete response rate, and the secondary endpoint was to explore the association between clinical outcomes and genetic polymorphisms CYP2A6 (*4, *7, *9 and *10), UMPS G638C, and three ABCB1 genotypes (C1236T, C3435T, and G2677T). ResultsNinety-one patients were given study treatment, and 90 underwent surgery. Pathologic complete response was noted in 10 patients (11.1%). There was no grade 4 or 5 toxicity; 20 (22.0%) experienced grade 3 toxicities, including diarrhea (10, 11.0%), abdominal pain (2, 2.2%), and anemia (2, 2.2%). Relapse-free survival and overall survival at 5 years were 88.6% and 94.2%, respectively. Patients with the UMPS 638 CC genotype experienced significantly more frequent grade 2 or 3 diarrhea (p for trend = 0.018). Conclusions
Preoperative chemoradiation with tegafur-uracil 400 mg/m2/day with leucovorin was feasible, but did not meet the expected pathologic complete response rate. The UMPS 638 CC genotype might be a candidate biomarker predicting toxicity in patients receiving tegafur-uracil/leucovorin-based preoperative chemoradiation for locally advanced rectal cancer. Trial registrationISRCTN11812525, registered on 25 July 2016. Retrospectively registered.
... Simultaneously, these cells acquired a mesenchymal phenotype, with enhanced expression of N-cadherin, vimentin, Notch-1, and morphology changes, consistent with EMT induction. 165 In addition to U87, other human glioma cell lines, resistant to BCNU (carmustine) showed alterations related with EMT, such as fibroblast-morphology, enhanced migratory and invasive capacity, and decreased E-cadherin expression, with concomitant enhanced expression of vimentin and MMP-9. 166 It is known that CSC is considered to be more resistant to toxic injuries and chemoradiation therapy than differentiated cells. ...
Glioblastoma multiforme (GBM) is the most aggressive form of brain cancer due to its highly invasive nature that impedes the surgical removal of all tumor cells, making relapse inevitable. However, the mechanisms used by glioma cells to invade the surrounding tissue are still unclear. In this context, epithelial-to-mesenchymal transition (EMT) has emerged as a key regulator of this invasive state and although the real relevance of this program in malignant glioma is still controversial, it has been strongly associated with GBM malignancy. EMT is a very complex process regulated by several families of transcriptional factors through many signaling pathways that form a network that allows cancer cells to acquire invasive properties and penetrate the neighboring stroma, resulting in the formation of an advantageous microenvironment for cancer progression and metastasis. In this systematic review, we focus on the molecular mechanisms of EMT including EMT-factors, drug resistance, miRNA, and new therapeutic strategies. In addition, we address controversial questions about mesenchymal shift in GBMs with a bioinformatics analysis to show that in terms of epithelial and mesenchymal phenotype, the majority of GBMs samples analyzed have a profile more mesenchymal than epithelial. If induced, this phenotype can be shifted toward an even more mesenchymal phenotype in an EMT-like process in glioma cells. A better understanding of the molecular regulation of the EMT during tumor spreading will help to provide potential therapeutic interventions to target this program when treating GBM.
... ABCB1 is an ATP-dependent drug efflux transporter for a huge variety of substrates [35]. Although capecitabine has not been clearly identified as one of them, ABCB1 expression has been associated with resistance to 5-FU in modified cell lines [36]. The authors showed associations between the haplotype ABCB1*1 and an increased probability of reducing the dose of capecitabine, delaying initiation of therapy, or withdrawing the drug altogether, as well as of diarrhea and overall toxicity. ...
Adverse reactions to capecitabine-based chemotherapy limit full administration of cytotoxic agents. Likewise, genetic variations associated with capecitabine-related adverse reactions are associated with controversial results and a low predictive value. Thus, more evidence on the role of these variations is needed. We evaluated the association between nine polymorphisms in MTHFR, CDA, TYMS, ABCB1, and ENOSF1 and adverse reactions, dose reductions, treatment delays, and overall toxicity in 239 colorectal cancer patients treated with capecitabine-based regimens. The ABCB1*1 haplotype was associated with a high risk of delay in administration or reduction in the dose of capecitabine, diarrhea, and overall toxicity. CDA rs2072671 A was associated with a high risk of overall toxicity. TYMS rs45445694 was associated with a high risk of delay in administration or reduction in the dose of capecitabine, HFS >1 and HFS >2. Finally, ENOSF1 rs2612091 was associated with HFS >1, but was a poorer predictor than TYMS rs45445694. A score based on ABCB1-CDA polymorphisms efficiently predicts patients at high risk of severe overall toxicity (PPV, 54%; sensitivity, 43%) in colorectal cancer patients treated with regimens containing capecitabine. Polymorphisms in ABCB1, CDA, ENOSF1,and TYMS could help to predict specific and overall severe adverse reactions to capecitabine.
