靶向survivin基因siRNA在食管癌细胞增殖和凋亡中作用的研究

Study of the effect of siRNA targeted against survivin on cell proliferation and apoptosis of esophageal cancer cells

目的研究survivin基因特异性RNA干扰对食管癌Eca-109细胞株体外增殖能力和凋亡的影响。方法构建靶向survivin的小干扰RNA(siRNA)表达载体,用脂质体法转染Eca-109细胞后,采用半定量RT-PCR、Western Blot检测转染前后survivin mRNA及蛋白表达水平的改变,采用噻唑兰(MTT)法检测细胞的生长增殖情况,采用流式细胞术测定细胞的凋亡情况。结果靶向survivin的序列特异性的siRNA可以有效地抑制Eca-109细胞survivin基因的表达。转染靶向survivin的siRNA表达质粒可以显著抑制Eca-109细胞的增殖(细胞接种24 h、48 h后增殖抑制率分别为21.05%和33.96%),诱导明显的细胞凋亡[转染24 h、48 h后的凋亡率分别为(8.03±1.05)%和(6.22±1.06)%]。结论survivin基因特异性RNA干扰可以显著抑制食管癌Eca-109细胞的体外增殖并诱导明显的细胞凋亡。

Objective To investigate the effect of siRNA targeted against survivin on cell proliferation and apoptosis of Eca-109 esophageal cancer cells. Methods A siRNA plasmid expression vector against survivin was constructed and transfected into Eca-109 cells with LipofectamineTM2000. The changes of survivin mRNA and protein expression were detected by semi-quantitive RT-PCR and Western Blot. Cell proliferation was detected by MTT assay. Cell apoptosis was evaluated by flow cytometry. Results The sequence-specific siRNA effectively suppressed survivin expressing at both mRNA and protein levels. Survivin expression surpression significantly inhibited the proliferation （ at 24 h and 48 h after cell seeding, the proliferation inhibition rate was 21.05% and 33.96% respectively） and induced apoptosis of Eca-109 cells （ at 24 h and 48 h after transfection, apoptosis rate was （ 8. 03 ± 1.05 ） % and （6. 22 ± 1.06） % respectively. Conclusions siRNA targeted against survivin can effectively suppress Eca-109 cell proliferation and significantly induce Eea-109 cell apoptosis.