采用RNA干扰技术沉默survivin基因对口腔鳞状细胞癌细胞凋亡的影响

Effects of survivin gene silencing by small interfering RNA on apoptosis of oral squamous cell carcinoma

目的应用RNA干扰(RNAi)技术抑制口腔鳞状细胞癌细胞株HSC-3细胞凋亡抑制因子survivin基因的表达,促进HSC-3细胞的凋亡。方法合成针对survivin基因的小干扰RNA(siRNA),转染对数生长期的HSC-3细胞。采用半定量逆转录聚合酶链反应检测HSC-3细胞中survivin mRNA的表达,MTT法测定细胞活性,tunnel分析和流式细胞术检测HSC-3细胞的凋亡,并设阴性对照组和空白对照组。结果转染siRNA组survivin mRNA的表达明显低于阴性对照组和空白对照组;MTT法测定细胞活性,阴性对照组和空白对照组没有明显差别,但siRNA组的细胞活性明显下降;tunnel分析显示,转染siRNA组的凋亡细胞明显多于阴性对照组和空白对照组;流式细胞术分析,转染siRNA组的凋亡率(24.99%±1.33%)明显高于阴性对照组(1.24%±0.13%)和空白对照组(0.10%±0.02%)。结论采用siRNA沉默survivin基因能促进口腔鳞状细胞癌细胞的凋亡,survivin siRNA基因治疗有可能成为口腔鳞状细胞癌治疗的新技术。

Objective To apply the small interfering RNA （siRNA） targeting survivin to inhibit expression of sur- vivin gene in HSC-3 oral squamous cell carcinoma and to increase the apoptosis of HSC-3 cells. Methods siRNA was synthesized and transfected into oral squamous cell carcinoma HSC-3 cells. Compared with negative control group and blank control group, semi-quantitive reverse transcription-polymerase chain reaction （RT-PCR） was used to detect survivin mRNA. Cellular viability was detected by MTT and HSC-3 cell apoptosis was determined by tunnel and flow cytometry. Results Expression of survivin mRNA was decreased significantly in siRNA group compared with negative control group and blank control group. Cellular viability was not affected in negative control group and blank control group, but cellular viability in siRNA group was significantly decreased. As to the cellular apoptosis rate, the trans- leered group（24.99%±1.33%） was significantly higher than negative control group（1.24%±0.13%） and blank control group （0.10%±0.02%）. Conclusion Survivin gene silenced by siRNA might promote apoptosis of oral squamous cell carcinoma. Survivin siRNA gene therapy would become a new target point for oral squamous cell carcinoma.