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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10μmol/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。
Objective: To investigate the inhibitory effect of total-sulfated anti-telomerase antisense oligonucleotide (PS-ODN) on colon cancer LS-174T cells and to explore the mechanism of inhibition of colon cancer cell by telomerase inhibitor PS-ODN . Methods: The antisense technique was used to block the telomerase reverse transcription level to induce apoptosis. The inhibitory effect was evaluated by electron microscopy, FCM and PCR-Elisa. Results: The optimal dosage of PS-ODN was 10μmol / L. After 10 days, the colony formation rate of LS-174T cells in PS-ODN group was significantly lower than that in CPS-ODN group and control group (P <0.01) (P <0.01). The apoptosis peak appeared in PS-ODN group after 72h, but not in control group. The telomerase activity of PS-ODN group was significantly lower than that of CPS-ODN group and control group (P <0.01) ). Conclusion: The specific nucleotide sequence can inhibit telomerase activity, induce apoptosis, and lay a solid foundation for new approaches and new ideas for the treatment of tumors.