枸杞多糖增敏紫杉醇对人卵巢癌HO-8910PM细胞凋亡的影响和机制研究

Effects of lycium barbarum polysaccharide on the apoptosis of human ovarian cancer cell line HO-8910PM and its mechanism

目的探索枸杞多糖能否作为增敏剂协同增强紫杉醇对人上皮性卵巢癌HO-8910PM细胞凋亡和增殖的影响,并探讨其可能的作用机制。方法用CCK8法检测LBP、PTX单药作用细胞的半数抑制率,选取25%、50%半数抑制率浓度作为实验浓度;以流式细胞术检测细胞周期、细胞凋亡,RT-PCR和Western blot分别检测LBP、PTX协同作用后HO-8910PM细胞Survivin、Caspase-3、Bax、Bcl-2 mRNA和蛋白的表达。结果协同作用处理后,LBP协同PTX作用HO-8910细胞可见凋亡率大于单药,增益作用显著(P<0.05);LBP协同PTX作用HO-8910细胞可见G0/GI期、G2/M期细胞比例增加,对细胞周期抑制的增益作用显著;LBP协同PTX作用HO-8910细胞后可见Survivin、Bcl-2基因mRNA de蛋白表达显著减少(P<0.05),Caspase-3、Bax基因mRNA和蛋白表达量显著增多(P<0.05),均呈现浓度依赖性。结论枸杞多糖可作为协同增敏紫杉醇对HO-8910PM细胞的抗癌能力,其作用机制可能与Survivin、Bcl-2基因表达的下调,Caspase-3、Bax基因的表达上调有关。

Objective To investigate the effect of lycium barbarum polysaccharide as a reversal agent on the proliferation and ap- optosis of human ovarian epithelial carcinoma cell line HO8910PM and its possible mechanism. Methods Half inhibition rate of LBP and PTX single drug action cells was detected by CCK8 method. 25% and 50% of inhibition rate concentration were selected as experimental concentration. Cell cycle and apoptosis were detected by flow cytometry. The expressions of HO8910PM cells Survivin, caspase -3, Bax, bel - 2 mRNA and protein were detected by RT - PCR and western blot after the synergistic effects of LBP and PTX. Results After synergis- tic treatment, LBP synergistic PTX action on HO8910 cells showed that apoptosis rate was greater than the sum of single drug action, and the gain effect was significant （P 〈0.05）. LBP in cooperation with PTX in HO8910 cells showed that the proportion of cells at G0/GI stage and G2/M stage increased significantly ,and the cell cycle inhibition was significantly enhanced. LBP PTX action of HO8910 cells mediated by Survivin, BCL -2 gene mRNA DE protein expression was significantly reduced （P 〈 0.05 ）, Caspase 3, Bax gene mRNA and protein ex- pression significantly increased （P 〈 0.05 ）, and presented the concentration dependence. Conclusion LBP can be used as a synergistic sensitizing strong paclitaxel in the anti -cancer ability of HO8910PM cells,and its mechanism may be related to the down -regulation of Survivin ,bel -2 gene expression and the up- regulation of caspase- 3 and Bax gene expression.