绿原酸对肾癌A498和769-P细胞增殖、迁移和侵袭的影响

Effects of chlorogenic acid on the proliferation,migration and invasion of renal carcinoma A498 and 769-P cells

目的探讨绿原酸对肾癌A498和769-P细胞增殖、迁移和侵袭的影响以及其可能的分子机制。方法取人肾癌A498或769-P细胞培养,分为对照组和绿原酸(2μL,1μmol/L)组,培养72 h。采用MTT法检测细胞增殖能力;Transwell小室法检测细胞侵袭能力;划痕实验检测细胞迁移能力;ELISA法检测细胞上清液中IL-1β水平;qRT-PCR法检测细胞EPAS-1表达;Western blot检测细胞AKT/P65信号通路相关蛋白表达。结果绿原酸能抑制胃癌A498或769-P细胞增殖、侵袭和迁移的能力,降低细胞上清液中IL-1β水平。qRT-PCR和Western blot检测结果显示,与对照组比较,加入anti-IL-1β后,肾癌A498或769-P细胞中EPAS-1 mRNA及p-AKT、p-P65蛋白表达水平降低;与对照组比较,绿原酸组肾癌A498或769-P细胞中EPAS-1 mRNA及p-AKT、p-P65蛋白表达水平降低(P<0.05)。结论绿原酸能抑制肾癌细胞增殖、侵袭及迁移,其机制可能与抑制细胞IL-1β的分泌,从而抑制AKT/P65/EPAS-1通路有关。

Objective To investigate the effects of chlorogenic acid on the proliferation,migration and invasion of renal carcinoma A498 and 769-P cells and the possible molecular mechanism.Methods Human renal carcinoma A498 and 769-P cells were divided into control group and chlorogenic acid group(2μL,1μmol/L)and cultured for 72 h.The cell proliferation,invasion and migration were detected with MTT assay,Transwell assay and scratch test,respectively.The expressions of IL-1β,EPAS-1 and AKT/P65 signaling pathway related proteins were detected with ELISA,qRT-PCR and Western blot,respectively.Results Chlorogenic acid inhibited the proliferation,invasion and migration of renal carcinoma A498 and 769-P cells,and reduced the IL-1βlevel in the cell supernatant.Anti-IL-1βreduced the protein and mRNA expressions of EPAS-1,p-AKT and p-P65.Compared with the control group,the chlorogenic acid group had reduced mRNA and protein expressions of EPAS-1,p-AKT and p-P65(P<0.05).Conclusion Chlorogenic acid can inhibit the invasion and metastasis of renal carcinoma cells,and its mechanism may be related to inhibiting the secretion of IL-1β,thereby inhibiting the AKT/P65/EPAS-1 pathway.