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为了探讨MKI67在肝癌细胞发生发展中的作用,采用实时定量PCR方法检测人肝细胞癌QGY-7703细胞中MKI67基因表达水平,以及MKI67在肝细胞癌组织和癌旁正常组织中的表达情况,设计并合成针对MKI67的siRNA,利用脂质体转染法将其转入QGY-7703细胞内,通过MTT和细胞集落形成实验观察MKI67-siRNA对QGY-7703细胞生长活性和增殖能力的影响.实时定量PCR结果表明,MKI67在肝细胞癌组织中的表达水平明显高于癌旁正常组织(P<0.01).MTT和细胞集落形成实验结果显示,转染MKI67-siRNA的QGY-7703细胞生长活性和集落形成率明显低于对照组(P<0.01).由此得出结论:MKI67在肝癌细胞系QGY-7703细胞中的表达水平较高,且它在肝癌组织中的表达水平明显上调.同时,MKI67-siRNA可以有效抑制QGY-7703细胞的生长活性和增殖能力,提示MKI67可能与肝细胞癌的发生、发展相关.
To investigate the role of MKI67 in the development and progression of hepatoma cells, the expression of MKI67 in human hepatocellular carcinoma QGY-7703 cells and the expression of MKI67 in hepatocellular carcinoma tissues and adjacent normal tissues were detected by real-time PCR. The siRNA against MKI67 was synthesized and transfected into QGY-7703 cells by lipofectamine 2000. The effect of MKI67-siRNA on the growth activity and proliferation of QGY-7703 cells was observed by MTT assay and colony formation assay. The results of PCR showed that the expression level of MKI67 in hepatocellular carcinoma was significantly higher than that in adjacent normal tissues (P <0.01) .MTT and colony formation assay showed that the growth of MKI67-siRNA-expressing QGY-7703 cells and colonies The formation rate of MKI67 was significantly lower than that of the control group (P <0.01) .Therefore, it was concluded that MKI67 was highly expressed in hepatocellular carcinoma cell line QGY-7703 and its expression was significantly up-regulated in hepatocellular carcinoma.MKI67 -siRNA can effectively inhibit the growth activity and proliferation ability of QGY-7703 cells, suggesting that MKI67 may be associated with the occurrence and development of hepatocellular carcinoma.