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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。
OBJECTIVE: To investigate the inhibitory effect of ethyl acetate extract (Squilla Oratoria, ESO) and cisplatin (DDP) on the proliferation of human hepatoma HepG2 cells and its mechanism. Methods: MTT assay was used to determine the inhibitory effect of ESO and DDP alone or in combination on the proliferation of HepG2 cells. The combined effect index (CI) was calculated. The effect of ESO on cell cycle and apoptosis was detected by flow cytometry. HepG2 cells Survivin, Bax and Bcl-2 protein expression levels. To establish a subcutaneous HepG2 cell xenograft model in nude mice and observe the inhibitory effect of ESO and DDP alone or in combination on the transplanted tumor. Results: ESO effectively inhibited the growth of HepG2 cells in a concentration-dependent manner, promoted cell apoptosis and blocked the cell cycle in S phase. When combined with DDP, the inhibition of HepG2 cells proliferation showed a synergistic effect. Mortality increased significantly (P <0.05). ESO downregulated Survivin and Bcl-2 protein expression, up-regulated Bax protein expression in a concentration-dependent manner (P <0.05). Tumor inhibition test in nude mice found that, with the increase of ESO concentration, the rate of tumor growth slows down, and the inhibition effect of combination group is more obvious. Conclusion: ESO can inhibit the proliferation of HepG2 cells through the cell cycle arrest and apoptosis. Combined with cisplatin can inhibit the proliferation of HepG2 cells.