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目的 研究反义寡聚核苷酸 (asONs)能否抑制人膀胱肿瘤T2 4细胞系端粒酶活性和增殖活性。方法 设计并合成 2条针对端粒酶RNA模板区的asONs片段 ,在其作用于T2 4细胞第 5天时 ,用 7复孔法用聚合酶链反应 酶标法 (PCR ELISA )检测其对端粒酶活性的影响 ;分别在实验的第 1、3、5、7天 ,用 7复孔四唑盐比色试验 (MTT)法检测其对细胞增殖活性的影响。结果 经不同浓度 (0、5、10 μmol/L)处理 5d后T2 4细胞端粒酶活性受到抑制 ,吸光度 (A )值分别为0 .79± 0 .10、0 .2 4± 0 .14 ;经 10 μmol/L处理 1、3、5、7d ,T2 4细胞的增殖活性逐渐下降 ,A值分别为 0 .5 9± 0 .11、0 .3 9± 0 .0 9、0 .2 1± 0 .0 2、0 .19± 0 .0 7,各组分别比较差异有非常显著性(P <0 .0 0 1)。结论 asONs能够同时抑制人膀胱肿瘤T2 4细胞系端粒酶及细胞增殖活性 ,提示asONs可能通过抑制端粒酶活性达到抑制人膀胱肿瘤T2 4细胞系的增殖
Objective To investigate whether antisense oligonucleotides (asONs) can inhibit the telomerase activity and proliferative activity of human bladder tumor T2 4 cell line. Methods Two asONs fragments targeting the telomerase RNA template region were designed and synthesized. On day 5 of T2 4 cells, they were tested for their resistance to telomere by polymerase chain reaction (ELISA) Enzyme activity. The effects of cell proliferation on the 1st, 3rd, 5th and 7th day of the experiment were detected by MTT assay. Results The telomerase activity of T2 4 cells was inhibited after treated with different concentrations (0, 5, and 10 μmol / L) for 5 days. The absorbance values (A) were 0.79 ± 0.10 and 0.24 ± 0.14 ; The proliferative activity of T2 4 cells gradually decreased after treated with 10 μmol / L for 1, 3, 5 and 7 days, and the A values were 0.59 ± 0.11, 0.39 ± 0.90, 1 ± 0. 0 2,0 .19 ± 0 .0 7, the differences between the groups were significant (P <0. 001). Conclusion asONs can simultaneously inhibit telomerase activity and cell proliferation in human bladder tumor T2 4 cell line, suggesting that asONs may inhibit the proliferation of human bladder tumor T2 4 cell line by inhibiting telomerase activity