阿托伐他汀对人胃癌AGS细胞增殖、自噬和糖代谢的影响及机制

Effect and mechanism of atorvastatin on the proliferation,autophagy and glucose metabolism of AGS cells in human gastric cancer

目的探究阿托伐他汀对人胃癌AGS细胞增殖、自噬和糖代谢的影响及机制。方法通过预实验考察低、中、高浓度(12.5、25、50μmol/L)阿托伐他汀对AGS细胞活力的影响,筛选作用浓度。正式实验分为对照组(不做干预)、阿托伐他汀组(25μmol/L)、阳性对照组(50 mg/L 5-氟尿嘧啶)、抑制剂组[25μmol/L阿托伐他汀+10μmol/L磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶B(Akt)信号通路抑制剂LY294002]和激活剂组(25μmol/L阿托伐他汀+10μmol/L PI3K/Akt信号通路激活剂SC79),干预24 h,检测细胞糖代谢情况(葡萄糖和乳酸含量)和细胞增殖率,以及细胞中自噬相关蛋白轻链3(LC3)Ⅰ、LC3Ⅱ及PI3K/Akt信号通路相关蛋白的表达。结果中、高浓度阿托伐他汀均能显著抑制AGS细胞活力(P<0.05),正式实验选择25μmol/L阿托伐他汀进行后续实验。与对照组相比,阳性对照组和阿托伐他汀组细胞中葡萄糖和乳酸含量、细胞增殖率以及p-PI3K/PI3K、p-Akt/Akt比值均显著降低(P<0.05),LC3Ⅰ、LC3Ⅱ蛋白表达水平均显著升高(P<0.05)。与阿托伐他汀组相比,抑制剂能显著加强上述指标变化(P<0.05),激活剂能显著逆转上述指标变化(P<0.05)。结论阿托伐他汀可抑制人胃癌AGS细胞的糖代谢和增殖,促进细胞自噬,其作用机制可能与抑制PI3K/Akt信号通路相关。

OBJECTIVE To explore the effects and mechanism of atorvastatin on the proliferation,autophagy and glucose metabolism of AGS cells in human gastric cancer.METHODS The effects of low,medium and high concentrations of atorvastatin(12.5,25,50μmol/L)on the viability of AGS cells were investigated through preliminary experiments,and the concentration of action was screened.The formal experiment was divided into control group(no intervention),atorvastatin group(25μmol/L),positive control group(50 mg/L 5-fluorouracil),inhibitor group[25μmol/L atorvastatin+10μmol/L phosphatidylinositol-3-kinase(PI3K)/protein kinase B(Akt)signaling pathway inhibitor LY294002]and activator group(25μmol/L atorvastatin+10μmol/L PI3K/Akt signaling pathway activator SC79),all of which were treated for 24 h.Glucose metabolism(glucose and lactic acid contents)and cell proliferation rate were detected,as well as the expression of autophagy-associated protein light chain 3(LC3)Ⅰ,LC3Ⅱand PI3K/Akt signaling pathway-associated proteins in cells.RESULTS Both medium and high concentrations of atorvastatin could significantly inhibit the viability of AGS cells(P<0.05),and 25μmol/L atorvastatin was selected for the official experiment for follow-up experiments.Compared with the control group,the contents of glucose and lactic acid,cell proliferation rate,p-PI3K/PI3K and p-Akt/Akt ratios in the positive control group and atorvastatin group were significantly decreased(P<0.05),and the protein expression levels of LC3Ⅰand LC3Ⅱwere significantly increased(P<0.05).Compared with the atorvastatin group,the inhibitor further promoted the changes in the above indexes(P<0.05),and the activator significantly reversed the changes in the above indexes(P<0.05).CONCLUSIONS Atorvastatin could inhibit glucose metabolism and proliferation of AGS cells in human gastric cancer and promote autophagy.The mechanism may be related to the inhibition of the PI3K/Akt signaling pathway.