小白菊内酯调控自噬抑制食管癌细胞增殖和转移的作用研究

Study on the effect of parthenolide regulating autophagy on the inhibition of the proliferation and metastasis of esophageal cancer cells

目的:研究小白菊内酯(PTL)调控自噬抑制食管癌细胞增殖和转移的作用机制。方法:常规培养食管癌细胞Eca109,采用MTS法检测不同浓度PTL对Eca109细胞增殖能力的影响;采用自噬诱导剂Rapamycin或自噬抑制剂3-MA处理细胞,分为NC组、PTL组、PTL+Rapamycin组和PTL+3-MA组;Transwell实验检测细胞迁移能力;裸鼠皮下成瘤实验检测食管癌细胞体内生长能力;western blotting检测细胞中自噬相关蛋白P62、Beclin1和LC3Ⅱ蛋白表达。结果:PTL呈浓度依赖性地抑制食管癌细胞增殖(P<0.05)。与NC组比较,PTL组细胞迁移能力及裸鼠成瘤能力降低(均P<0.05)。与NC组比较,PTL组P62、Beclin1和LC3Ⅱ蛋白表达量升高(P<0.05);与PTL组比较,PTL+Rapamycin组P62、Beclin1和LC3Ⅱ蛋白表达量升高,PTL+3-MA组P62、Beclin1和LC3Ⅱ蛋白表达量降低(均P<0.05)。结论:PTL通过促进自噬抑制食管癌细胞增殖和迁移及裸鼠体内移植瘤生长。

Objective:To study the mechanism of parthenolide(PTL) inhibiting the proliferation and metastasis of esophageal cancer cells by regulating autophagy.Methods:The esophageal cancer cells Eca109 were conventionally cultured,and the MTS experiment was used to detect the effect of different concentrations of PTL on the proliferation of Eca109 cells.The cells were treated with autophagy inducer Rapamycin or autophagy inhibitor 3-MA and divided into NC group,PTL group,PTL+Rapamycin(autophagy activator) group,and PTL+3-MA(autophagy inhibitor) group.Transwell assay was used to detect the migration of cells.The growth of esophageal cancer cells in vivo was evaluated by using subcutaneous tumorigenesis in nude mice.Western blotting was used to detect autophagy-related proteins such as P62,Beclin1 and LC3 II expressions.Results:PTL inhibited the proliferation of Eca109 cells in a concentration-dependent manner(P<0.05).Compared with the NC group,the migration of cells,tumorigenicity in nude mice were significantly decreased in PTL group(P<0.05).Compared with NC group,the protein expressions of P62,Beclin1 and LC3 II in PTL group were increased(P<0.05).Compared with PTL group,the protein expression levels of P62,Beclin1 and LC3 II in PTL+Rapamycin group were increased while decreased in PTL+3-MA group(P<0.05).Conclusion:PTL inhibit the proliferation and migration of esophageal cancer cells and the growth of transplanted tumor in nude mice by promoting autophagy.