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Archive > Volume 23 Issue 6 > 2016,23(6):766-772. DOI:10.3872/j.issn.1007-385X.2016.06.005 Prev Next
Basic Research
Mechanism of hypoxia-inducible factor 1-α to participate epithelial mesenchymal transition and DNA homologous recombination repair of colorectal cancer cells
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Abstract:
Objective:To investigate effect of hypoxic microenviroment in vitro which built by cobalt chloride (CoCl2) on epithelial mesenchymal transition and DNA homologous recombination repair of colorectal cancer SW480 and Caco-2 line cells and to explore their mechanisms. Methods: After the SW480 and Caco-2 cells were treated with various concentrations of CoCl2 for 72 h, proliferation abilities of the cells were detected by CCK8 assay, migration and invasion abilities of the cells detected by scratch test and transwell assay, cell cycles and apoptosis status of the cells detected by flow cytometry (FCM), changes of mRNA and protein levels of associated gene in the cells detected by RT-PCR and Western blotting assays. Results: CCK8 assay prompted that proliferation ability of the cells significantly increased after hypoxia (P<0.05); scratch and transwell tests suggested that under hypoxia, migration and invasion abilities of the cells obviously enhanced (P<0.05); FCM disclosed that after anoxic treatment the cells were maintained at S phage and apoptosis rates of the cells significantly decreased (P<0.05); RT-PCR assay showed that mRNA levels of HIF-1α and RAD51 in the cells after hypoxic treatment raised (P<0.05); Western blotting assay indicated that under hypoxic microenviroment in the cells expression of HIF1-α enhanced (P<0.05), expression of epithelial mesenchymal transition (EMT) associated proteins, namely E-cadherin was down-regulated, expressions of vimentin and transcription inhibitor (snail) were up-regulated (P<0.05), expressions of DNA homologous recombination associated protein BRCA1 and RAD51 were up-regulated (P<0.05), phosphorylation levels of down stream key molecule Akt1 on PI3K/AKT (phosphatidyl inositol-3-hydroxy kinase) signaling pathway and ATM kinase encoded by ataxia-telangiectasia mutant gene were significantly enhanced (P<0.05). Conclusion: Chronic hypoxia enviroment could promote abilities of proliferation, migration and invasion of the colorectal cancer SW480 and Caco-2 cells as well as inhibit apoptosis of the cells. Mechanism of the actions might related to HIF-1α/PI3K and HIF-1α/ATM signaling pathways mediated EMT and DNA homologous recombination repair process.
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Project Supported:
Project supported by the Natural Science Foundation of Chongqing Municipal Science and Technology Commission (No. Cstc2012jjA10038)
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