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目的探讨冬凌草甲素联合survivin反义核苷酸(反义链)对前列腺癌PC-3细胞株增殖和凋亡以及survivin m RNA和蛋白的影响。方法常规培养PC-3细胞,用四甲基偶氮唑盐法(MTT法)检测survivin反义链联合冬凌草甲素对PC-3细胞增殖的影响;流式细胞仪(FCM)检测PC-3细胞凋亡率;以Calcu Syn药效学软件计算联合指数(CI)评价survivin反义链联合凌草甲素对PC-3细胞的联合效应,并通过荧光定量PCR和Western blot方法检测PC-3细胞survivin基因和蛋白表达变化。结果 survivin反义链转染PC-3细胞后,可以显著抑制PC-3细胞增殖,且能诱导PC-3细胞凋亡;冬凌草甲素联合survivin反义链对PC-3细胞增殖抑制率较两者单用组明显增强,显示出协同效应(Fa<0.8);冬凌草甲素和survivin反义链联用对PC-3细胞凋亡诱导作用更为显著(P<0.01);荧光定量PCR及Western blot显示反义链和冬凌草甲素均使survivin m RNA和蛋白表达下降,两者联合使survivin m RNA和蛋白表达下降更明显(P<0.01)。结论 survivin基因反义链和冬凌草甲素均能明显抑制PC-3增殖、诱导细胞凋亡和下调survivin基因和蛋白表达;两者联合作用有协同效应。
Objective To investigate the effects of oridonin combined with survivin antisense nucleotide (antisense strand) on the proliferation and apoptosis of prostate cancer cell line PC-3 and the expression of survivin m RNA and protein. Methods PC-3 cells were cultured routinely. The effects of antisense survivin and oridonin on the proliferation of PC-3 cells were detected by MTT assay. The cytotoxicity of PC-3 cells by flow cytometry (FCM) -3 cell apoptosis rate; Calcu Syn pharmacodynamics software to calculate the joint index (CI) evaluation of survivin antisense chain combined with the formidonin on PC-3 cells combined effect, and by fluorescence quantitative PCR and Western blot detection of PC -3 cells survivin gene and protein expression changes. Results The antisense siRNA transfected PC-3 cells could significantly inhibit the proliferation of PC-3 cells and induce the apoptosis of PC-3 cells. The antitumor activity of oridonin plus survivin inhibited the proliferation of PC-3 cells (Fa <0.8). The combination of oridonin and survivin antisense was more effective in inducing the apoptosis of PC-3 cells (P <0.01), and the fluorescence Quantitative PCR and Western blot showed that both antisense RNA and Oridonin decreased the expression of survivin m RNA and protein, and the combination of them decreased the expression of survivin m RNA and protein more significantly (P <0.01). Conclusion Both the antisense strand of survivin gene and oridonin can significantly inhibit the proliferation of PC-3 cells, induce the apoptosis of cells and down-regulate the expression of survivin gene and protein. The combined effect of them has a synergistic effect.