ESO联合DDP对肝癌HepG2细胞增殖的抑制效应及其机制

目的： 探讨海洋生物口虾咕乙酸乙酯提取物（ethyl acetate extract of Squilla Oratoria , ESO）联合顺铂（cisplatin，DDP）对人肝癌HepG2细胞增殖的抑制效应及其机制。 方法： MTT法测定不同浓度ESO和DDP 单独或联合处理对HepG2细胞增殖的抑制作用，并计算联合效应指数（CI）；流式细胞术检测ESO对细胞周期和细胞凋亡的影响，Western blotting检测HepG2细胞Survivin、Bax和Bcl-2的蛋白表达水平。建立裸鼠皮下HepG2细胞移植瘤模型，并观察ESO和DDP单用或联用对移植瘤的抑制效应。 结果： ESO能有效地抑制HepG2细胞的生长（呈浓度依赖性），促进细胞凋亡，将细胞周期阻滞在S期；与DDP联合用药时，对HepG2细胞增殖的抑制呈现出协同效应，凋亡率明显增加（ P <0.05）。ESO可下调Survivin和Ｂcl-2蛋白表达、上调Bax蛋白表达，且呈浓度依赖性（ P <0.05）。裸鼠成瘤抑制实验发现，随着ESO浓度的增大，肿瘤增长的速度减慢，以联合用药组抑制作用更为明显。 结论： ESO可通过细胞周期阻滞及促细胞凋亡作用抑制HepG2细胞增殖；与顺铂联合用药，能对HepG2细胞增殖产生协同抑制效应。

Anti-hepatoma effect of ethyl acetate extract of Squilla oratoria (ESO) in combination with cisplatin

Objective : To evaluate the anti-hepatoma effect of ethyl acetate extract of Squilla oratoria (ESO) in combination with cisplatin (DDP) in vitro and in vivo . Methods: Human hepatoma HepG2 cells were treated with different concentrations of ESO alone or in combination with DDP. After 24 h of treatment, cell viability was determined by MTT assay, cell cycle and apoptosis were assessed by flow cytometry, and the protein content of survivin, Bcl-2 and Bax was determined by Western blotting analysis. For in vivo assessment, HepG2 cells were transplanted to nude mice and animals with confirmed tumor development one week after transplantation were left untreated (control) or treated with ESO and DDP, either each alone or two in combination, for two weeks before sacrifice and measurement of tumor number and volume. Results: ESO effectively inhibited HepG2 cell proliferation ( P <0 05), induced S phase arrest ( P <0 05) and apoptosis ( P <0.05), increased the protein content of Bax ( P <0.05) and decreased the protein content of survivin and Bcl-2 ( P <0.05) in a dose-dependent manner in vitro . All these effects of ESO were synergistically enhanced by DDP ( P <0.05). In vivo , ESO robustly reduced the number and volume of hepatoma tumors in nude mice transplanted with HepG2 cells in a dose-dependent manner ( P <0 01) and cisplatin significantly synergized this effect of ESO. Conclusion: ESO may have an anti-hepatoma activity and this activity can be synergized by cisplatin.