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目的 :观察反义端粒酶RNA对SMMC772 1细胞的作用 ,并检测 p16、p2 1的表达情况 ,探讨端粒酶在肝癌发生过程中的可能作用及反义端粒酶RNA在肝癌治疗中的意义。方法 :经脂质体介导的方法将 pBBS2 12 hTR(反义端粒酶RNA真核表达载体 )导入SMMC772 1细胞中 ,细胞克隆转移扩大培养后 ,采用流式细胞仪、TRAP PCR ELISA、电镜、免疫组化技术结合图像分析 ,在检测SMMC772 1/ pBBS2 12 hTR细胞的端粒酶活性、凋亡的同时 ,检测p16、p2 1表达情况。 结果 :与对照组比较 ,反义端粒酶RNA在抑制SMMC772 1细胞端粒酶活性、诱发细胞发生凋亡的同时 ,p16、p2 1蛋白表达水平显著增高。结论 :转染反义端粒酶RNA在封闭细胞端粒酶活性表达、促进SMMC772 1细胞凋亡的同时 ,伴发 p2 1与p16表达水平增高。对研究肝癌发病机制及治疗有益
OBJECTIVE: To observe the effect of antisense telomerase RNA on SMMC772 1 cells and to detect the expression of p16 and p21, and to explore the possible role of telomerase in the development of hepatocellular carcinoma and the antisense telomerase RNA in hepatocellular carcinoma significance. METHODS: The pBBS2 12 hTR (antisense telomerase RNA eukaryotic expression vector) was introduced into SMMC772 1 cells by liposome-mediated method. After the cells were expanded and cultured, the growth of the cells was analyzed by flow cytometry, TRAP PCR ELISA and electron microscopy Immunohistochemistry and image analysis were used to detect the expression of p16 and p21 while detecting the telomerase activity and apoptosis of SMMC772 1 / pBBS2 12 hTR cells. Results: Compared with the control group, the antisense telomerase RNA inhibited the telomerase activity of SMMC772 1 cells and induced the apoptosis of cells, meanwhile, the expression of p16 and p21 proteins increased significantly. CONCLUSION: The transfection of antisense telomerase RNA can enhance the expression of telomerase activity in the closed cells and promote the apoptosis of SMMC772 1 cells. To study the pathogenesis and treatment of liver cancer