DOWN-REGULATED CD147 EXPRESSION BY RNA INTERFERENCE TO INHIBIT HT29 CELLS PROLIFERATION,INVASION AND

来源 :江苏省生物化学与分子生物学第九届会员代表大会暨第十次学术会议 | 被引量 : 0次 | 上传用户:qaz123_family
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  Objective: In this study, we aimed to reduce the expression of CD147 by RNA silencing, investigating the effect of CD 147 silencing to the human colorectal cancer cells proliferation, invasion and drug sensitivity.Methods: We constructed a novel short hairpin RNA (shRNA) expression vector pYr-mir30-shRNA.The CD147 shRNA plasmid was transferred to human colorectal cancer cell line HT29.The expression of CD147, MCT1 and MCT4 were monitored by quantitative real-time RT-PCR and Western blotting, respectively.By gelatin zymography assay determined the MMP-2 and MMP-9 activities.The intracellular lactaten concentration was determined by the lactic acid assay kit.WST-8 assay was applied to determine the HT29 cells proliferation and the chemosensitivity to cisplatin.Invasion assay was used to determine the invasion of HT29 cells.In addition, we selected the 4-6-week-old female nude mice to establish subcutaneous tumor model of colorectal cancer, and applie-d the immunohistochemical analysis to detect CD147 expression in vivo.Results: The results showed that the CD147 and MCT1 expression were significantly reduced at both mRNA and protein levels, as well as the MMP-2 and MMP-9 activities.The proliferation and invasion ability were also decreased in HT29 cells, but increased chemosensitivity to cisplatin.In vivo, the CD 147 expression was also significantly decreased, and reduced the tumor growth after CD147 gene silencing.Conclusions: The results demonstrated that silencing of CD147 expression inhibited the proliferation and invasion of colorectal cancer cells, but increased its chemosensitivity to cisplatin and decreased the secretion of MMPs in colorectal cancer cells, suggesting CD 147 silencing might be an adjuvant gene therapy strategy to chemotherapy.
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