白藜芦醇调控磷脂酰肌醇3-激酶/蛋白激酶B通路对结肠癌细胞增殖凋亡的影响

Effects of resveratrol on proliferation and apoptosis of colon cancer cells by regulating PI3K/Akt pathway

目的探究白藜芦醇调控磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)通路对结肠癌细胞增殖、凋亡的影响。方法体外培养人结肠癌SW480细胞,噻唑蓝(MTT)法检测人结肠癌SW480细胞增殖,将细胞分成对照组、低剂量白藜芦醇组(20μmol/L)、中剂量白藜芦醇组(40μmol/L)及高剂量白藜芦醇组(80μmol/L)。细胞划痕实验法测定人结肠癌SW480细胞迁移能力,Transwell小室检测人结肠癌SW480细胞侵袭能力,流式细胞仪检测人结肠癌SW480细胞凋亡,吖啶橙染色法检测人结肠癌SW480细胞自噬,实时定量聚合酶链反应(RT-PCR)法检测各组细胞B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关蛋白(Bax)、半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)mRNA水平,蛋白质免疫印迹法检测各组细胞PI3K-Akt信号通路。结果与对照组比较,20、40和80μmol/L白藜芦醇导致细胞存活率降低(P<0.05),且不同浓度白藜芦醇培养48 h最为适宜。与对照组对比,白藜芦醇不同剂量组中,细胞迁移能力减弱(P<0.05),且细胞的迁移率随着白藜芦醇浓度的提高逐渐减少。在对照组的基础上经白藜芦醇处理的实验组细胞凋亡率增加(P<0.05)。细胞凋亡率随白藜芦醇浓度增加而逐步提高(P<0.05)。在实验组中,与对照组比较,细胞在接受白藜芦醇处理后细胞自噬数量增加,且随着白藜芦醇浓度的提高,细胞自噬数量逐步增加(P<0.05)。相较于对照组,各剂量的白藜芦醇组Bcl-2下降、Bax与Caspase-3增加。相较于对照组,不同剂量白藜芦醇组显示出PI3K和p-Akt蛋白下降;不同剂量白藜芦醇组Akt蛋白差异无统计学意义(P>0.05)。结论白藜芦醇可通过介导PI3K/Akt信号通路促进结肠癌细胞凋亡,降低其增殖能力,减弱细胞迁移和侵袭能力,以控制结肠癌的侵袭和恶化。

Objective To explore the effects of resveratrol on the proliferation and apoptosis of colon cancer cells by regulating the PI3K/Akt pathway.Methods Human colon cancer SW480 cells were cultured in vitro and the proliferative capacity was evaluated using the MTT assay.The cells were divided into a control group,a low-dose resveratrol group(20μmol/L),a medium dose resveratrol group(40μmol/L),and a high-dose resveratrol group(80μmol/L).The migration abilities of the cells were assessed by the scratch assay,while their invasion capabilities were determined using a Transwell chamber.Apoptotic responses were quantified through flow cytometry and acridine orange staining.Furthermore,the mRNA expression levels of Bcl-2,Bax,and Cas-pase-3 were analyzed by RT-PCR.Additionally,the activity protein of the PI3K-Akt signaling pathway was in-vesti-gated in each group utilizing the Western.Results Compared with the control group,20,40,and 80μmol/L resveratrol significantly reduced cell survival rate(P<0.05),and 48 h of culture at different concentrations of resveratrol was most suitable.Moreover,the cell migration ability was significantly weakened in different doses of resveratrol groups(P<0.05),and the cell migration rate gradually decreased with the increase of resveratrol con-centration.On the basis of the control group,the experimental group cells treated with resveratrol showed a sig-nifi-cant increase in apoptosis rate(P<0.05).In addition,in this study,we observed a positive correlation between cell apoptosis rate and resveratrol concentration,and the cell apoptosis rate gradually increased with the increase of resveratrol concentration(P<0.05).Compared with the control group,the experimental group cells showed a sig-nifi-cant increase in autophagy after treatment with resveratrol,and the autophagy quantity gradually increased with the increase of resveratrol concentration(P<0.05).Compared with the control group,human colon cancer SW480 cells treated with different doses of resveratrol showed a significant decrease in Bcl-2 level,while a signifi-cant in-crease in Bax and caspase-3 levels.Compared with the control group,the various doses of resveratrol sig-nificantly decreased PI3K and p-Akt protein levels.However,there was no significant difference in Akt protein between dif-ferent doses of resveratrol groups(P>0.05).Conclusion Resveratrol can promote apoptosis of colon cancer cells by mediating the PI3K/Akt signaling pathway,reduce their prolifera-tion ability,weaken cell migra-tion and inva-sion ability,and control the invasion and deterioration of colon cancer.