α-倒捻子素通过加剧自噬影响胃癌细胞的增殖和凋亡

Alpha-mangostin affects the proliferation and apoptosis of gastric cancer cells by increasing autophagy

目的探讨α-倒捻子素对人胃癌细胞增殖和凋亡的影响及其分子机制。方法CCK8法检测α-倒捻子素对SGC7901细胞活力的影响,免疫荧光和流式细胞术检测α-倒捻子素对SGC7901细胞凋亡及细胞周期的影响,Western blot法检测细胞凋亡及自噬相关蛋白的表达情况。统计分析的正态性检验采用Shapir-Wilk;服从正态分布的多组间比较采用单因素方差分析;方差不齐时,采用Welch矫正检验,两两间比较采用Games-Howell检验。结果 CCK8法结果显示,不同浓度的α-倒捻子素(10、15、20、25、30μmol/L)处理后组间细胞活力的差异有统计学意义(P<0.05)。qPCR结果表明,LC3而非Caspase蛋白的mRNA水平与α-倒捻子素浓度呈正相关(r=0.976,P<0.05)。在15μmol/L而非10μmol/L的α-倒捻子素处理系统中,6-氨基-3-甲基嘌呤(3-MA,10μmol/L)、巴弗洛霉素A(10μmol/L)、LY294002(10μmol/L)等自噬抑制剂均能显著缓解α-倒捻子素对SGC7901细胞的杀伤作用(P<0.05)。结论α-倒捻子素可能主要通过诱导自噬性死亡而影响人胃癌细胞的活力。

Objective To investigate the possible molecular mechanism for alpha(α)-mangostin's inhibition of the proliferation and apoptosis of human gastric cancer cells.Methods Human gastric adenocarcinoma SGC7901 cell line was treated withα-mangostin.CCK8 method was used to measure the viability of SGC7901 cells.The effect ofα-mangostin on apoptosis and cell cycle was determined by immune fluorescence and flow cytometry.The expression of the relevant proteins was detected using Western blot.The shapiro-wilk test was performed for evaluation of deviation from normality.Normally distributed data was analyzed with one-way ANOVA.Welch test was used in data with heterogeneity of variance and multiple compared by Games-Howell test after that.ResultsCCK8 results showed that cell viability differed significantly among groups treated with different concentrations ofα-mangostin(10,15,20,25,and 30μmol/L)(P<0.05).QPCR data showed that the concentration ofα-mangostin was positively correlated with mRNA level of LC3 but not caspase protein(r=0.976,P<0.05).In15μmol/L but not 10μmol/Lα-mangostin treatment system,the autophagy inhibitors 3-MA(10μmol/L),bafilomycin A(10μmol/L)and LY294002(10μmol/L)could significantly alleviateα-mangostin's killing effect on SGC7901 cells(P<0.05).Conclusion The anti-tumor effects ofα-mangostin against human gastric adenocarcinoma cells in vitro can be partly attributed to apoptosis,autophagy and arresting cell phase.