南蛇藤提取物联合顺铂用药对人肝癌HepG2细胞的协同作用

The synergy effects of Celastrus extract combine with cisplatin in HepG2 Cells

目的观察南蛇藤提取物(Celastrus orbiculatus extract,COE)联合顺铂(Cisplatin,DDP),对人肝癌Hep G2细胞增殖、侵袭迁移和凋亡的协同作用。方法将对数期Hep G2细胞,分为对照组(不加药)、COE组(40 mg·L^(-1))、顺铂组(DDP 2 mg·L^(-1))及联合组(同时加入40 mg·L^(-1)的COE和2 mg·L^(-1)的DDP)。药物作用24 h后,MTT法计算细胞增殖抑制率,划痕实验及Transwell实验分析细胞的侵袭迁移,流式细胞术检测细胞凋亡率,Western blot法分析上皮间质转化相关蛋白(E-cadherin、N-cadherin和vimentin)以及凋亡相关蛋白(Bax和Bcl-2)的表达水平。结果与对照组相比,各组药物均能抑制肝癌细胞的增殖,缩短迁移距离,促进细胞凋亡(P<0.05)。Western blot结果表明,与对照组相比,各组药物均能抑制人肝癌Hep G2细胞内N-cadherin、vimentin及Bcl-2蛋白的表达水平,同时增加E-cadherin及Bax蛋白的表达量。与单一用药相比,COE联合DDP用药,作用效果更显著(P<0.05)。结论南蛇藤提取物可协同顺铂,促进肝癌细胞凋亡,抑制肝癌细胞侵袭迁移,其分子机制可能与上皮间质转化及Bcl-2/Bax通路有关。

Objective To investigate the synergy effects of C. orbiculatus extracts （COE） combined with cisplatin （DDP） on proliferation, invasion, migration and apoptosis in HepG2 cells. Methods The HepG2 cells were treated with different drugs （40 mg ·L^-1COE, 2 mg ·L^-1DDP, 40 mg ·L^-1COE combine with 2 mg ·L^-1DDP） for 24 h. The cell proliferation inhibition rate was calculated by MTT. The cell migration was observed by cell scratch test and Transwell Assay. The apoptosis rate was measured by using flow cytometry. The protein expression levels of E - cadherin, N - cadherin, vimentin and Bcl - 2 were detected by Western blotting. Results Compared with the control, each drug can significantly inhibit the invasion and migration. Meanwhile, each drug promotes the apoptosis of HepG2 cells. The protein expression of N - cadherin, vimentin and Bcl - 2 were decreased significantly. Meanwhile, the protein expression of E - cadherin and Bax were significant enhanced. Compared with the single drug, the synergy effects have been shown obviously while COE combine with DDP （P 〈 0.05 ）. Conclusion There is a synergy effects between COE and cisplatin in HepG2 cells. The molecular mechanisms may be related to the EMT process and Bcl - 2/Bax pathway.