靶向survivin的siRNA诱导胰腺癌细胞凋亡的实验研究

目的 采用RNA干扰技术（siRNA）阻断survivin基因的表达，观察其抑制胰腺癌细胞增殖及诱导胰腺癌细胞凋亡的作用。方法 构建靶向survivin的siRNA质粒表达载体，采用Lipofectamine^TM2000转染胰腺癌细胞PC-2，采用半定量RT-PCR、免疫组化技术检测转染前后PC-2细胞survivin基因表达的变化；采用MTT法检测对PC-2细胞增殖的抑制作用：采用流式细胞术检测其诱导PC-2细胞凋亡的作用。结果 靶向survivin的序列特异性的siRNA可以高效地抑制PC-2细胞survivin基因表达，在mRNA水平其表达抑制率为81．25％，在蛋白质水平其表达抑制率为74．24％。转染靶向survivin的siRNA质粒表达载体可以显著抑制PC-2细胞的增殖，细胞接种24、48h后其增殖抑制率分别为28．00％和33.38％：转染后24、48h可以诱导8．46％、7．53％的细胞凋亡。结论 所构建的靶向survivin的siRNA质粒表达载体可以有效地阻断PC-2细胞survivin基因表达。阻断survivin基因表达可以显著地抑制PC-2细胞的增殖并在一定程度上诱导其凋亡，靶向survivin的siRNA在胰腺癌的基因治疗中具有一定的价值。

siRNA targeted against survivin induces apoptosis of pancreatic cancer cells

Objective To investigate the effect of a sequence-specific small interfering RNA (siRNA) in suppressing survivin expression and cell proliferation and inducing apoptosis of PC-2 cells. Methods The plasmid expression vector of siRNA targeted against survivin was constructed and transfected into PC-2 cells with Lipofectamine?000. The changes of survivin expression were detected by semi-quantitative RT-PCR and immunohistochemical SP methods. The effect of siRNA in suppressing the proliferation of PC-2 cells was detected by MTT assay, and its role in inducing PC-2 cell apoptosis evaluated by flow cytometry. Results The sequence-specific siRNA effectively suppressed survivin expression at both mRNA and protein levels with inhibition rate of 81.25% at mRNA level and 74.24% at protein level. Survivin expression suppression significantly inhibited the proliferation of PC-2 cells, and at 24 and 48 h after cell seeding, the proliferation inhibition rate was 28.00% and 33.38% respectively; 24, 48 h after the transfection, apoptosis occurred in 8.46% and 7.53% of the cells, respectively. Conclusions The plasmid expression vector for the siRNA against survivin constructed in the study can effectively and specifically suppress survivin expression in PC-2 cells, and blocking survivin expression suppresses PC-2 cell proliferation and induces cell apoptosis. siRNA targeted against survivin has a potential value in gene therapy for pancreatic cancer.