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目的 观察反义端粒酶 RNA对 SMMC- 772 1肝癌细胞系的作用 ,探讨抗端粒酶治疗在原发性肝癌治疗中的意义 .方法 采用脂质体介导的方式将反义端粒酶 RNA真核表达载体 p BBS2 12 / h TR转入 SMMC772 1细胞中 ,经潮霉素筛选 ,克隆细胞株扩增鉴定后 ,采用 TRAP- PCR- EL ISA,FCM及电镜检测反义端粒酶 RNA对转染细胞端粒酶活性和细胞凋亡的影响 ;同时进行裸鼠荷瘤生长实验观察 .结果 TRAP- PCR- EL ISA法检测细胞端粒酶活性的结果为 :实验组吸光度值为 0 .980 ,对照组吸光度值为 2 .86 1;FCM检测细胞凋亡结果为 :实验组 13.8% ,对照组未见有凋亡峰形成 ,电镜观察发现转染细胞表现出典型的凋亡细胞形态 .裸鼠移植瘤生长实验发现 ,转染细胞生长明显受到抑制作用 ,说明反义端粒酶 RNA显著抑制 SMMC772 1细胞的端粒酶活性 ,并且对其有显著的促凋亡作用 .结论 反义端粒酶 RNA能有效地抑制肝癌细胞端粒酶活性 ,同时显著促进癌细胞凋亡 .
Objective To observe the effect of antisense telomerase RNA on SMMC-772 1 hepatocellular carcinoma cell line and explore the significance of anti-telomerase therapy in the treatment of primary hepatocellular carcinoma. Methods Liposome-mediated antisense telomerase The RNA eukaryotic expression vector p BBS2 12 / h TR was transfected into SMMC772 1 cells. After selection by hygromycin, the cloned cell lines were amplified and identified. The antisense telomerase RNA was detected by TRAP-PCR-EL ISA, FCM and electron microscope. The effect of telomerase activity and apoptosis in transfected cells was observed simultaneously. The results of TRAP-PCR-EL ISA assay showed that the absorbance of the experimental group was 0. 980, the absorbance value of the control group was 2.86 1; the result of apoptosis detected by FCM was: 13.8% of the experimental group, and no apoptotic peak was formed in the control group. The electron microscope showed that the transfected cells showed typical apoptotic cell morphology. Growth experiments in nude mice transplanted tumors showed that the growth of transfected cells was significantly inhibited, indicating that antisense telomerase RNA significantly inhibited the telomerase activity of SMMC7721 cells, and had a significant proapoptotic effect on them. Conclusion Antisense end Granzyme RNA can effectively inhibit Hepatoma telomerase activity, while significantly promote apoptosis of cancer cells.