端粒酶反义寡核苷酸体外抑制宫颈癌细胞端粒酶活性的研究

Inhibition of telomerase antisense oligonucleotides on telomerase activity of cervical carcinoma cell in vitro

目的探讨端粒酶反义寡核苷酸(ASON)对宫颈癌Hela细胞端粒酶活性的影响。方法针对端粒酶RNA的ASON作用于Hela细胞,同时以加有转染剂FuGENE6者为对照组,计数连续作用1,3,5 d后的细胞数。TRAP方法测定端粒酶的活性。结果ASON组与随机引物及空白对照组比较,端粒酶活性降低,细胞的增殖速度延缓;有转染剂FuGENE6的ASON组的细胞增殖数目和端粒酶活性显著低于无FuGENE6的ASON组;细胞对有FuGENE6的ASON的摄取显著多于单纯的ASON。在FuGENE6的介导下,ASON对端粒酶活性的抑制与ASON的浓度和治疗时间呈正相关。结论针对端粒酶RNA的ASON能抑制Hela细胞的端粒酶活性,转染剂FuGENE6可以增强端粒酶ASON对宫颈癌细胞端粒酶活性的抑制作用。

Objectives: To investigate the effects of telomerase antisense oligonucleotides on telomerase activity of cervical carcinoma Hela cells. Methods: Hela cells were treated by antisense oligonucleotides targeted to the RNA temp late region of telomerase and oligonucleotide with FuGENE6 was observed as controls. Number of the cells was counted from 1 to 5 days. The telomerase aitivity was performed using PCR-TRAP. Results: The ASON reduced the telomerase activity and number of cells compared with the random primer and the empty controls. The ASON with FuGENE6 inhibited the telomerase activity and Hela cells growth more efficiently than the ASON without FuGENE6 and the efficiency depended on the dose of ASON and the time of treatment. Conclusions: ASON targeted the RNA template region of telomerase inhibite the telomerase activity of Hela cells; Transfection regant FuGENE6 enhance the efficiency of inhibition.