HTR反义寡核苷酸抑制鼻咽癌细胞生长及对其致瘤能力的影响

Growth inhibition of nasopharyngeal carcinoma HNE-1 cell strain and effect on its tumorigenic ability by antisense oligodeoxynucleotide targeting telomerase RNA

目的:研究针对端粒酶RNA(Human telomerase RNA,HTR)的反义寡核苷酸(Antisense oligodeoxynucleotide,AS-ODN)抑制鼻咽癌HNE-1细胞株生长及对其致瘤能力的影响。方法:PCR-ELISA法检测细胞内端粒酶活性。MTT法及平板集落形成实验评价细胞增殖活性。电镜观察细胞超微结构的改变。将经AS-ODN处理的HNE-1细胞接种于裸鼠皮下,观察其致瘤情况。结果:HTR反义寡核苷酸作用HNE-1细胞后,细胞端粒酶活性受到明显抑制;细胞增殖受到明显抑制,此作用有剂量、时间依赖性和序列特异性;电镜发现少量细胞肿胀坏死,但未见细胞凋亡和大片坏死灶。AS-ODN降低了HNE-1细胞的致瘤能力,裸鼠成瘤时间延长,肿瘤生长速度变慢。结论:HTR反义寡核苷酸可通过抑制鼻咽癌HNE-1细胞端粒酶活性而抑制细胞的生长,并抑制其对裸鼠的致瘤能力。

Objective：To investigate the growth inhibition of nasopharyngeal carcinoma cell strain and effect on its tumorigenic ability by AS-ODN targeting telomerase RNA. Methods：Telomerase activity was analysed by PCR-ELISA technique. Proliferation of HNE-1 cells was examined by MTT test and clone formation test. Ultrastructure changes were examined by electron microscope. AS-ODN treated HNE-1 cells were implanted subcutaneously into nude mice to observe tumor growths. Resets：When HNE-1 cells were incubated with the AS-ODN targeting telomerase RNA,telomerase activities of HNE-1 cells were significantly inhibited;proliferation of HNE-1 cells was significantly inhibited,which showed a dose-dependent and timedependent correlation and was sequential specific;swelling or necrosis was found in a small proportion of HNE-1 cells by electron microscopy but no apoptosis and large necrosis area could be found. AS-ODN reduced the tumorigenic ability of HNE-1 cell strain. The tumor formation time was prolonged and tumor growth was slowed down. Conclusion：Through inhibiting the telomerase activity of HNE-1 cell strain,the AS-ODN targeting telomerase RNA can inhibit its growth and reduce its tumorigenic ability.