低剂量阿司匹林诱导产生的自噬减弱其对人肝癌HepG2细胞系的抑制作用

目的 研究一定浓度的阿司匹林对人肝癌HepG2细胞系的增殖抑制作用及产生抑制作用可能的原因.方法 采用MTT法和平板克隆实验研究阿司匹林对人肝癌HepG2细胞增殖活力的影响.吖啶橙荧光染色法观察阿司匹林对人肝癌HepG2细胞内自噬小体的影响.Western blot法检测腺苷酸活化蛋白激酶(AMPK)、哺乳动物雷帕霉素靶蛋白(mTOR)在人肝癌HepG2细胞中的表达.结果 10 mmol/L浓度的阿司匹林能抑制人肝癌HepG2细胞的增殖活力,但增加HepG2细胞中自噬小体的数量,增加AMPK表达,降低 mTOR的表达,且和40 μm自噬抑制剂氯喹(CQ)联合使用时,CQ能够增强10 mmol/L浓度阿司匹林对人肝癌HepG2细胞的增殖抑制作用.结论 联合自噬抑制剂CQ减弱浓度为10 mmol/L的阿司匹林诱导产生的自噬从而明显增强其抗HepG2作用.

Low-dose aspirin induced autophagy comprises its inhibitory effect on HepG2 cells

Objective To study the inhibitory effect of aspirin on proliferation of human hepatocellular carcinoma HepG2 cell line and its possible mechanismt. Methods MTT assay and plate cloning experiments was used to detect proliferation of human hepatoma HepG2 cells. Effects of aspirin on autophagosomes in HepG2 cells were detected by acridine orange fluorescence staining. The expression of adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) and mammalian target of rapamycin (mTOR) protein in human hepatocellular carcinoma HepG2 cells was detected by Western blot. Results 10 mmol/L concentration of aspirin could inhibit the proliferation of HepG2 cells, but increase the number of autophagosomes of HepG2 cells, increase AMPK expression, decrease mTOR expression. After combination treatemnt with 40 μmol/L autophagy inhibitor chloroquine (CQ), CQ could enhance the inhibitory effect of 10 mmol/L aspirin on proliferation of human hepatoma HepG2 cells. Conclusion Combination treatment with autophagy inhibitor CQ attenuates 10 mmol/L aspirin-induced autophagy thus enhance its anti-HepG2 effect.