脂质体介导的端粒酶反义寡核苷酸体外转染率及对人膀胱癌EJ细胞的影响

In vitro transfection efficiency of telomerase antisence oligodeoxy nucleic acids carried by liposome and its influence on human bladder cancer cell line EJ

目的通过以脂质体为载体介导端粒酶反义寡核苷酸(antisense oligodeoxynucleotide,asODN)转染人膀胱癌EJ细胞,促进反义寡核苷酸对膀胱癌EJ细胞的生长抑制。方法利用脂质体为载体将针对端粒酶RNA模板区的asODN转染膀胱癌EJ细胞;荧光显微镜观察细胞转染率;PCR-ELISA法测定端粒酶活性;MTT法检测反义寡核苷酸对膀胱癌细胞的抑制。结果脂质体介导的asODN在膀胱癌EJ细胞中的转染率1/2h、4h、8h分别为15%、56%、80%,并且细胞的端粒酶活性和细胞生长明显被抑制,与对照组比较,具有显著性差异(p(0.05)。结论脂质体介导的端粒酶asODN能够有效抑制膀胱癌EJ细胞端粒酶活性和生长,可应用于实验性肿瘤基因治疗研究和端粒酶与膀胱癌关系的进一步研究。

Objectives To improve inhibition of human bladder cancer cell line EJ through telomerase antisense oligedeoxynucleotide （esODN） mediated by liposome. Methods To investigate the transfection efficiency of esODN, telomerase antisense oligodeoxynucleotides with fluorescein 5/- isothiocyanate （5/- F1TC ） were mediated by liposome to human bladder cancer cell line EJ and fluorescence microscopy was used in fixed cells. Telonerase activation was measured through PCR - ELISA, and influence of telomerase esODN on inhibition of bladder cancer line EJ tested by MTT. Results In the presence of liposome, the transfection rate of esODN was 15% ,56%, 80% in 1/2h ,4h,8h respectively in bladder cancer line EJ. After 72h treated with esODN, telomerase activation and growth rate of tumor celia markedly decreased, and there was significance compared with control group（ p 〈 0.05 ）. Conclusions Specific esODN carded by cationic lipids could inhibit telomerase activity and growth of bladder cancer line EJ in vitro, it could be used in the research of cancer gene therapy and in the exploration of relationship between telomerase and neoplasm.