龙胆苦苷对人上皮性卵巢癌细胞增殖、凋亡、迁移的影响及机制

Effects of gentiopicroside on proliferation,apoptosis,and migration of ovarian cancer cells

目的观察龙胆苦苷(GPS)对人上皮性卵巢癌HO8910细胞增殖、凋亡及迁移能力的影响,并探讨其可能的作用机制。方法培养人上皮性卵巢癌HO8910细胞,随机分为对照组、GPS低剂量组、GPS中剂量组、GPS高剂量组。对照组单纯加入0. 01%DMSO培养48 h,GPS低、中、高剂量组分别在加入10、20与40μmol/L GPS的DMEM培养液中培养48 h。采用MTT法观察各组细胞增殖能力,计算细胞增殖抑制率;采用TUNEL实验观察各组细胞凋亡情况,计算细胞凋亡率;采用细胞划痕实验观察各组细胞迁移能力,计算细胞迁移率。采用Western blotting法检测p-P38有丝分裂原激活的蛋白激酶(MAPK)、促凋亡蛋白B细胞白血病淋巴瘤相关蛋白(Bax)、抗凋亡蛋白B淋巴细胞/白血病-2(Bcl-2)的表达。结果与对照组相比,GPS中剂量组、GPS高剂量组细胞增殖抑制率、迁移率均降低,凋亡率均升高(P <0. 05或0. 01)。与对照组相比,GPS中剂量组、GPS高剂量组细胞内Bax、p-P38MAPK表达上调,Bcl-2表达下调(P <0. 05或0. 01)。结论 GPS具有抑制人上皮性卵巢癌HO8910细胞增殖及迁移并促进其凋亡的作用,其机制可能与激活P38 MAPK信号通路相关。

Objective To investigate the effects of gentiopicroside (GPS) on the proliferation, apoptosis, and migration of ovarian cancer cells. Methods The ovarian cancer cells HO8910 were cultured in vitro, and then were divided into the control group, low-dose, medium-dose, and high-dose GPS groups. The cells in the drug groups were treated with 10, 20, and 40 μmol/L of GPS for 48 h, and the cells in the control group were only cultured in vitro with 0.01% DMSO for 48 h. MTT was used to detected the effect of GPS on proliferation of HO8910 cells;the apoptosis rate of HO8910 was detected by TUNEL;the cell scratch test was used to detected the effect of GPS on cell migration;and the phosphorylation level of P38 mitogen-activated protein kinase (p-P38MAPK) and the expression of mitochondrial apoptosis pathway proteins (Bax), B-lymphocyte/leukemia -2 (Bcl-2) protein were were detected by Western blotting. Results Compared with the control group, the proliferation inhibitory and migration rates of cells decreased significantly and the apoptosis rates increased in the medium-dose and high-dose GPS groups ( P <0.05 or P <0.01). Compared with the control group, the expression of Bax3 and p-P38MAPK was up-regulated, and the expression of Bcl-2 protein was down-regulated in the medium-dose and high-dose GPS groups ( P <0.05 or P <0.01). Conclusion The GPS can inhibit the proliferation and migration of ovarian cancer cell lines and promote its apoptosis by activating the P38MAPK pathway.