姜黄素对人肝癌SMMC-7721细胞AMPK信号通路机制及其凋亡影响的研究

Study on the mechanism of AMPK signaling pathway and its effect on apoptosis of human hepatocellular carcinoma SMMC-7721 cells by curcumin.

作者信息

Zhang Y-J, Xiang H, Liu J-S, Li D, Fang Z-Y, Zhang H

机构信息

Department of Interventional Radiology & Vascular Surgery, Hunan Provincial People's Hospital, Changsha, China.

出版信息

Eur Rev Med Pharmacol Sci. 2017 Mar;21(5):1144-1150.

PMID:28338175

Abstract

OBJECTIVE

Liver cancer is a common malignant tumor in the digestive system. Curcumin is a kind of phenolic pigment, which is extracted from herbage and has a plenty of physiological roles in anti-inflammation, anti-oxidation and anti-tumor. In our study, human hepatoma SMMC-7721 cell lines were selected and treated with curcumin to detect its effects on the apoptosis and AMPK signaling pathway.

MATERIALS AND METHODS

Human liver cancer cell strain SMMC-7721 was cultured and treated with different curcumin concentrations for different times followed by measuring the changes of cell proliferation activity and cycle by MTT and flow cytometry, respectively. Protein expression of Bcl-2, Bax and Caspase-3 were tested by Western blot, and the activation level of AMPK was also detected.

RESULTS

Different concentrations of curcumin could inhibit the proliferation of tumor cells in a dose-dependent manner. After 48 h inhibition by curcumin with a concentration of 40 mmol/L, the inhibitory effect was more obvious with statistically significant (p<0.05). The number of human liver cancer SMMC-7721 cells increased in G1 stage and decreased in S stage after treated with different concentrations of curcumin. During the G1 stage to the S stage, inhibition occurred and the effect of curcumin intervention group with 40 mmol/L was more evident than that of 10 mmol/L group, 20 mmol/L group and the control group with statistically significant (p<0.05). SMMC-7721 cell stains had been intervening by curcumin with concentrations of 10 mmol/L, 20 mmol/L and 40 mmol/L for 12 h, 24 h and 48 h, as the drug concentration increased, the reaction time prolonged, the protein expressions of Bcl-2 and Survivin were significantly decreased and Bax protein expression was significantly increased (p<0.05).

CONCLUSIONS

Curcumin decreased the proliferation activity of tumor cells, increased the cell quantities in G1 stage and decreased the cell numbers in S stage in human liver cancer SMMC-7721 cells. The Bcl-2 and Survivin proteins were downregulated and Bax protein was upregulated; furthermore, the AMPK signaling pathway was activated.

摘要

目的

肝癌是消化系统常见的恶性肿瘤。姜黄素是一种从草本植物中提取的酚类色素，在抗炎、抗氧化和抗肿瘤方面具有多种生理作用。在本研究中，选用人肝癌SMMC - 7721细胞系并用姜黄素处理，以检测其对细胞凋亡和AMPK信号通路的影响。

材料与方法

培养人肝癌细胞株SMMC - 7721，用不同浓度的姜黄素处理不同时间，然后分别通过MTT法和流式细胞术检测细胞增殖活性和细胞周期的变化。通过蛋白质免疫印迹法检测Bcl - 2、Bax和Caspase - 3的蛋白表达，同时检测AMPK的激活水平。

结果

不同浓度的姜黄素能以剂量依赖的方式抑制肿瘤细胞的增殖。用浓度为40 mmol/L的姜黄素作用48 h后，抑制作用更明显，差异具有统计学意义（p<0.05）。不同浓度姜黄素处理后人肝癌SMMC - 7721细胞G1期细胞数量增加，S期细胞数量减少。在从G1期到S期的过程中出现抑制，40 mmol/L姜黄素干预组的作用比10 mmol/L组、20 mmol/L组及对照组更明显，差异具有统计学意义（p<0.05）。用浓度为10 mmol/L、20 mmol/L和40 mmol/L的姜黄素分别干预SMMC - 7721细胞株12 h、24 h和48 h，随着药物浓度增加、作用时间延长，Bcl - 2和Survivin的蛋白表达显著降低，Bax蛋白表达显著增加（p<0.05）。