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目的:研究靶向survivin的(小分子干扰RNA)siRNA和(氟尿嘧啶)5-FU联用对肝癌细胞HepG2的增殖抑制及凋亡的影响。方法:将HepG2细胞分为空白对照组、阴性对照组、5-FU处理组、siRNA转染组、5-FU+siRNA转染组。转染采用脂质体法。RT-PCR法检测HepG2细胞survivin mRNA转录水平;MTT法检测靶向survivin的siRNA和5-FU对HepG2细胞增殖的抑制作用;流式细胞术检测HepG2细胞凋亡情况。结果:空白对照组、阴性对照组、5-FU处理组survivin mRNA表达无明显变化(P>0.05),siRNA转染组、5-FU+siRNA转染组survivin mRNA表达明显下降(F=280.326,q=4.72~7.34,P<0.05)。5-FU+siRNA转染组增殖抑制率为51.58%±1.35%,与其它各组相比抑制率明显增高(F=280.326,q=5.27~9.84,P<0.05)。5-FU+siRNA组与其它各组相比细胞凋亡率明显增高(F=13568.68,q=110.47～327.16,P<0.01)。结论:将靶向survivin的siRNA和5-FU联合应用可以显著抑制肝癌细胞survivin基因表达,并协同抑制HepG2细胞增殖,共同发挥诱导细胞凋亡作用。
OBJECTIVE: To investigate the effect of survivin-targeted (small interfering RNA) siRNA and (5-fluorouracil) 5-FU on the proliferation inhibition and apoptosis of HepG2 cells. Methods: HepG2 cells were divided into blank control group, negative control group, 5-FU treatment group, siRNA transfection group, and 5-FU+siRNA transfection group. Transfection using liposome method. The expression of survivin mRNA in HepG2 cells was detected by RT-PCR. The proliferation of HepG2 cells was detected by MTT assay and siRNA targeting survivin. The apoptosis of HepG2 cells was detected by flow cytometry. RESULTS: The expression of survivin mRNA in blank control group, negative control group and 5-FU group had no significant change (P>0.05). The expression of survivin mRNA in siRNA transfection group and 5-FU+siRNA transfection group was significantly decreased (F=280.326, q=4.72~7.34, P<0.05). The inhibition rate of 5-FU+siRNA transfection group was 51.58%±1.35%. The inhibition rate was significantly higher than other groups (F=280.326, q=5.27 to 9.84, P<0.05). The apoptosis rate of 5-FU+siRNA group was significantly higher than that of other groups (F=13568.68, q=110.47-327.16, P<0.01). Conclusion: The combined use of siRNA targeting survivin and 5-FU can significantly inhibit the expression of survivin gene in hepatocellular carcinoma cells and synergistically inhibit the proliferation of HepG2 cells, which together play a role in inducing apoptosis.