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目的探讨小干扰RNA(siRNA)对人肾癌细胞增殖基因Ki67表达及其增殖、凋亡的影响。方法将Ki67 siRNA(100 nmol/L)转染人肾癌786-0细胞。采用RT-PCR、免疫印迹、免疫组化技术检测Ki67 mRNA及蛋白表达,MTT法检测细胞增殖,免疫组化TUNEL法检测细胞凋亡。结果Ki67 siRNA处理组786-0细胞Ki67 mRNA表达(37．6±1．9)%、Ki67蛋白表达(46．4±0．9)%、免疫组化Ki67表达吸光度(A)值52．5±2．3,阴性siRNA对照组分别为(97．3±0．9)%、(95．3±0．9)%、114．5±4．9,2组比较差异均有统计学意义(P＜0．01)。Ki67 siRNA处理组细胞增殖抑制率(63．6±1．6)%、凋亡细胞阳性率(41．7±0．6)%,阴性siRNA对照组分别为(2．8±0．2)%、(10．3±1．4)%,2组比较差异有统计学意义(P＜0．01)。结论肿瘤增殖基因Ki67 siRNA能抑制人肾癌786-0细胞Ki67基因表达,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 the proliferation and apoptosis of human renal cell carcinoma cell line Ki67. Methods Ki67 siRNA (100 nmol / L) was transfected into human renal carcinoma 786-0 cells. The expression of Ki67 mRNA and protein was detected by RT-PCR, Western blotting and immunohistochemistry. Cell proliferation was detected by MTT assay and apoptosis by immunohistochemical TUNEL assay. Results Ki67 mRNA expression in Ki67 siRNA group was (37.6 ± 1.9)%, Ki67 protein expression was (46.4 ± 0.9)%, immunohistochemical Ki67 expression was 52.5 ± 2.3, the negative control group was (97.3 ± 0.9)%, (95.3 ± 0.9)%, 114.5 ± 4.9, the difference was statistically significant (P <0.01). Ki67 siRNA treatment group, the proliferation inhibition rate (63.6 ± 1.6)%, apoptotic cells (41.7 ± 0.6)%, negative siRNA control group were (2.8 ± 0.2) %, (10.3 ± 1.4)% respectively. The difference between the two groups was statistically significant (P <0.01). Conclusions Ki67 siRNA can inhibit the expression of Ki67 in human renal carcinoma 786-0 cells, and then inhibit its proliferation and apoptosis. It is expected that Ki67 siRNA will be an effective tool for gene therapy of renal carcinoma.