β-细辛醚对肾癌增殖及迁移侵袭的影响及其分子机制

Effects and mechanism ofβ-asarone on inhibiting cell proliferation/migration/invasion of renal cell carcinoma

目的研究β-细辛醚(β-As)对肾癌细胞的抑制作用及其分子机制,帮助阐明β-As抗肾肿瘤的作用并为其临床转化提供参考。方法本实验选用人肾癌细胞786-O和769-P进行体外实验,使用小鼠肾癌细胞Renca进行体内实验。应用MTT法、细胞划痕试验、细胞侵袭实验(Transwell)小室检测肾癌细胞体外增殖、迁移侵袭能力;应用蛋白印迹法检测自噬及上皮间质转化(EMT)等相关蛋白水平的变化;应用Balb/c小鼠皮下成瘤实验检测β-As在体内的抗肿瘤作用。结果β-As可通过剂量依赖的方式抑制肾癌细胞786-O和769-P体外增殖。β-As可上调E-cadherin的表达,下调N-cadherin、Vimentin的表达,并抑制其体外迁移侵袭能力。β-As可通过剂量依赖的方式上调LC3-Ⅱ、p-AMPK表达,下调p-mTOR、p-S6K的表达,而对总的AMPK、mTOR表达水平无显著影响。体内实验发现β-As可抑制肾癌细胞增殖。结论β-细辛醚通过自噬依赖性AMPK/mTOR信号通路对肾癌细胞的生长增殖、迁移侵袭及EMT进行抑制,在肾透明细胞癌尤其是进展期肾癌方面具有一定的临床转化潜能。

Objective To investigate the effects and molecular mechanism ofβ-asarone(β-As)on renal cell carcinoma(RCC).Methods Human RCC 786-O and 769-P cells were used in in vitro experiments,and murine RCC Renca cells were used in in vivo experiments.MTT,wound healing assay and Transwell assay were used to evaluate cell migration and invasion.Western blot was used to detect changes in protein levels during autophagy and epithelial-mesenchymal transition(EMT).Xenograft models of Balb/c mice were used to detect the anti-tumor effect ofβ-As in vivo.Resultsβ-As inhibited cell growth in a concentration-dependent manner;upregulated the expression of E-cadherin,but down-regulated the expressions of N-cadherin and Vimentin;inhibited cell proliferation,migration and invasion.β-As upregulated the expression of LC3 and p-AMPK,downregulated the expressions of p-mTOR and p-S6K in a concentration-dependent manner,but had no effects on the expressions of AMPK and mTOR.β-As inhibited the proliferation of RCC cells in vivo.Conclusionβ-As inhibits the proliferation,migration,invasion and EMT of RCC cells through the autophagy-dependent AMPK/mTOR signaling pathway,which may provide new ideas for the treatment of RCC,especially advanced RCC.