论文部分内容阅读
已知mi R-615-5p可抑制癌细胞的增殖,然而其具体分子机制尚不明确。本研究证明,mi R-615-5p通过负调节癌基因TRAF4,从而抑制NSCLC细胞的增殖。运用实时定量PCR检测NSCLC患者癌组织和癌旁正常组织、正常人肺支气管上皮细胞系HBE和3种人源NSCLC细胞系中mir-615-5p的表达,发现与正常的组织和细胞相比,mir-615-5p在NSCLC癌组织和癌细胞中表达水平显著降低;运用Western印迹检测HBE细胞和NSCLC细胞系中TRAF4蛋白的表达,发现TRAF4在NSCLC细胞中表达显著升高;MTT和CCK-8分析结果显示,转染mir-615-5p mimic可显著降低NSCLC细胞的增殖能力;生物学信息分析和萤光素酶报告基因检测结果显示,mir-615-5p可靶定结合TRAF4 m RNA,并下调TRAF4蛋白的水平;pc DNA-TRAF4转染后细胞增殖检测结果显示,过表达TRAF4能够消除mir-615-5p引起的细胞增殖抑制作用。综上所述,mir-615-5p通过靶定结合癌基因TRAF4的m RNA,下调TRAF4蛋白的水平,从而抑制NSCLC细胞的增殖。
It is known that mi R-615-5p can inhibit the proliferation of cancer cells, however, its specific molecular mechanism is not yet clear. This study demonstrates that mi R-615-5p inhibits the proliferation of NSCLC cells by negatively regulating the oncogene TRAF4. The real-time quantitative PCR was used to detect the expression of mir-615-5p in NSCLC patients with cancerous tissues and adjacent normal tissues, normal human bronchial epithelial cell line HBE and three human NSCLC cell lines. Compared with normal tissues and cells, The expression of TRAF4 protein in NSCLC cells was significantly increased by Western blotting. The expression of TRAF4 in NSCLC cells was significantly increased. The expressions of MTT and CCK-8 The results showed that the mir-615-5p transfected mir-615-5p mimic can significantly reduce the proliferation of NSCLC cells; biological information analysis and luciferase reporter gene test results show that mir-615-5p can target TRAF4 m RNA binding and Down-regulated the level of TRAF4 protein. The results of cell proliferation assay after transfection with pcDNA-TRAF4 showed that over-expression of TRAF4 could abolish the cell proliferation inhibition induced by mir-615-5p. Taken together, mir-615-5p inhibits the proliferation of NSCLC cells by targeting the mRNA that binds to the oncogene TRAF4, downregulating the level of TRAF4 protein.