【作者】 王锡山； 王贵玉； 徐海涛； 王宽； 刘明； 傅松滨； 耿敬姝； 张岂凡； 董新舒； 赵家宏；

【Author】 Wang Xi-Shan Wang Gui-Yu Xu Hai-Tao Wang Kuan Liu Ming Fu Song-Bin Geng Ye-Mei Zhang Qi-Fan Dong Xin-Shu Zhao Jia-Hong Department of Abdominal Surgery, The third hospital of Harbin Medical University Harbin 150001, P.R. China

【机构】 哈尔滨医科大学附属肿瘤医院；

【摘要】 目的:观察全硫化修饰的抗端粒酶反义寡核苷酸 (PS—ODN)对结肠癌LS-174T细胞的抑制作用,探讨端粒酶抑制剂PS—ODN对结肠癌细胞的抑制作用机制。方法: 采用反义技术在逆转录水平阻断端粒酶模板,从而诱导细胞凋亡,通过电镜、FCM、PCR—Elisa等方法评估抑制效果。结果:PS—ODN的最佳剂量为10 μmol／L,作用10天后, PS—ODN组LS—174T细胞集落形成率明显低于CPS—ODN 组和对照组(P<0．01),PS—ODN组细胞有明显的形态学改变,作用72h后PS—ODN组细胞出现凋亡峰,而对照组未出现,PS—ODN组细胞端粒酶活性明显低于CPS—ODN组和对照组相(P<0．01)。结论:特异的核苷酸序列可抑制端粒酶活性,诱导细胞凋亡,端粒酶介导的恶性肿瘤治疗策略是非常有前景的。恶性肿瘤中固有的人类端粒酶RNA的存在,使端粒酶成为PNA攻击的理想靶点。特定的核苷酸序列可抑制端粒酶活性,诱导细胞凋亡,为肿瘤的治疗提出新手段和新思路奠定了实验基础。更多还原

【Abstract】 Objective: To study the inhibitive effect of phosphorothioate anti-sense oligodeoxynucleotides (PSODN) on colorectal cancer LS-174T cells and the mechanism of telomerase inhibitor. Methods: PSODN was used to infect LS-174T cells and block human telomerase RNA (hTR) through anti-sense technology, which could induce LS-174T cells to apoptosis as result. The inhibitive effect of PSODN is evaluated by means of electron microscope (EMS), polymerase chain reaction-enzyme-linked immunosorbent assay (PCR-ELISA) and FCM assay. Results: PSODN showed a dose-time dependent inhibition of cell proliferation. The optimal dosage of PSODN was 10 lllllllll mol/L. After 10 days, PSODN group exhibited significantly different from control groups (P < 0.01). PCR-ELISA results indicated that telomerase activity of the PSODN group was obviously inhibited (P < 0.01). Meanwhile, the change on cellular number and morphology was observed. The cells in PSODN group showed apoptosis peak 72 hours later, but this didn’ t happen in control group. Conclusion: Specific sequence oligonucleotides can inhibit telomerase activity and lead cells to wither and die. The treatment strategy of malignant tumours induced by telomerase is much potential.更多还原