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目的探讨小干扰RNA(siRNA)对人肾癌细胞端粒酶基因表达、活性的抑制作用及对肾癌细胞增殖、凋亡的影响。方法将针对人端粒酶RNA(hTR)模板区的siRNA(100nmol/L)转染肾癌7860细胞,采用RTPCR检测7860细胞端粒酶mRNA表达,端粒重复序列扩增酶联免疫吸附法检测端粒酶活性,MTT法检测细胞增殖,免疫细胞化学TUNEL法检测细胞凋亡。结果hTRsiRNA处理组7860细胞端粒酶mRNA表达(40.7±1.5)%降低,端粒酶活性(32.7±2.3)%降低,细胞增殖抑制率(63.6±1.6)%增加,凋亡细胞阳性率(39.4±0.6)%增加。分别与阴性siRNA对照组比较差异有统计学意义(P<0.01)。结论siRNA能抑制人肾癌细胞端粒酶mRNA表达及活性,进而抑制其增殖、促进其凋亡,有望成为肾癌基因治疗的有效工具。
Objective To investigate the effect of small interfering RNA (siRNA) on telomerase gene expression and activity in human renal cell carcinoma and its effect on proliferation and apoptosis of renal cell carcinoma. METHODS: Human renal cancer 7860 cells were transfected with siRNA against human telomerase RNA (hTR) template (100 nmol / L). The expression of telomerase mRNA in 7860 cells was detected by RTPCR and telomerase reverse transcriptase-linked immunosorbent assay The activity of telomerase was detected by MTT assay. Cell apoptosis was detected by TUNEL assay. Results Telomerase activity (78.7 ± 1.5)%, telomerase activity (32.7 ± 2.3)%, cell proliferation inhibition rate (63.6 ± 1.6)% in hTRsiRNA treatment group increased significantly ± 0.6)% increase. Respectively, compared with negative siRNA control group, the difference was statistically significant (P <0.01). Conclusion siRNA can inhibit the expression and activity of telomerase mRNA in human renal cell carcinoma cells, and then inhibit its proliferation and apoptosis, which is expected to be an effective tool for gene therapy of renal cell carcinoma.