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- 2018
α-倒捻子素通过加剧自噬影响胃癌细胞的增殖和凋亡
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Abstract:
摘要：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。
ABSTRACT: 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 with α-mangostin. CCK8 method was used to measure the viability of SGC7901 cells. The effect of α-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. Results CCK8 results showed that cell viability differed significantly among groups treated with different concentrations of α-mangostin (10, 15, 20, 25, and 30μmol/L) (P<0.05). QPCR data showed that the concentration of α-mangostin was positively correlated with mRNA level of LC3 but not caspase protein (r=0.976, P<0.05). In 15μmol/L but not 10μmol/L α-mangostin treatment system, the autophagy inhibitors 3-MA (10μmol/L), bafilomycin A (10μmol/L) and LY294002 (10μmol/L) could significantly alleviate α-mangostin？？s killing effect on SGC7901 cells (P<0.05). Conclusion The anti-tumor effects of α-mangostin against human gastric adenocarcinoma cells in vitro can be partly attributed to apoptosis, autophagy and arresting cell phase
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