环吡酮胺抑制人胃癌AGS细胞生长、侵袭及其机制研究

Study of ciclopirox olamine inhibiting human AGS gastric cancer cells growth,invasion and the underlying mechanism

目的探讨环吡酮胺(ciclopirox olamine,CPX)抑制人胃癌AGS细胞生长、侵袭的作用及其机制。方法使用不同浓度CPX作用于人胃癌AGS细胞,采用CCK-8法明确CPX对AGS细胞体外增殖的影响,描绘CPX药物浓度-抑制率曲线图,计算其半数抑制浓度(IC_(50));采用细胞侵袭小室法(Transwell)检测CPX对人胃癌AGS细胞迁移、侵袭能力的影响;应用流式细胞仪检测CPX对人胃癌AGS细胞凋亡和细胞周期的影响;使用Western blotting法检测肿瘤相关迁移蛋白E-cadherin、CDK2、Bcl-2、Bax蛋白的表达情况。结果CPX能够抑制人胃癌AGS细胞的体外增殖活性,且呈现出明显的浓度及时间依赖性;IC_(50)值约13.6μmol/L,表明CPX在较小剂量时即对人胃癌AGS细胞产生毒性作用,且毒性作用的大小与药物浓度成正比。CPX能够明显抑制人胃癌AGS细胞的迁移、侵袭能力,且呈浓度依赖效应。CPX能够诱导人胃癌AGS细胞凋亡,且具有浓度依赖效应,当处理浓度较大时(20μmol/L),CPX可以阻滞人胃癌AGS细胞的细胞周期,这可能与阻滞G_(0)/G_(1)期有关。CPX可使人胃癌AGS细胞E-cadherin表达上调,从而抑制肿瘤细胞迁移;上调促凋亡因子Bax的表达,下调抗凋亡因子Bcl-2的表达,从而诱导人胃癌AGS细胞凋亡;下调CDK2的表达,从而使细胞周期阻滞在G_(0)/G_(1)期。结论CPX具有一定的抑制人胃癌AGS细胞增殖和迁移的能力,有望成为治疗胃癌的新型药物。

Objective To investigate the effects and molecular mechanism of ciclopirox olamine(CPX)inhibiting the growth,invasion of human AGS gastric cancer cells.Methods After different concentrations of CPX were applied to human AGS gastric cancer cells,the CCK-8 method was used to detect the growth inhibition of CPX on human gastric cancer AGS cells,and the curve of CPX drug concentration versus inhibition rate was drawn to calculate the half inhibitory concentration(IC_(50)).Transwell experiments were preformed to detect the effect of CPX on invasion and migration of gastric cancer AGS cells.The effects of CPX on the apoptosis and cell cycle of gastric cancer AGS cells were detected by flow cytometry.The expressions of E-cadherin,CDK2,Bcl-2 and Bax proteins were detected by Western blotting.Results CPX inhibited the in vitro proliferation of gastric cancer AGS cells in a concentration-and time-dependent manner;The IC_(50) value was about 13.6μmol/L,indicating that CPX exerted toxic effects on gastric cancer AGS cells at low dose,and the magnitude of the toxic effects was in direct proportion to the drug concentration.CPX significantly inhibited the migration and invasion of gastric cancer AGS cells in a concentration-dependent manner.CPX induced apoptosis of gastric cancer AGS cells in a concentration-dependent manner.When the concentration was relatively high(20μmol/L),CPX could organize the cell cycle of gastric cancer AGS,which might be related to the blockade of G_(0)/G_(1) phase.CPX up-regulated the expression of E-cadherin in gastric cancer AGS cells,and inhibited tumor cell migration.Up-regulated the expression of pro-apoptotic factor Bax and down-regulated the expression of anti-apoptotic factor Bcl-2 to induce apoptosis of gastric cancer AGS cells.The expression level of CDK2 was down-regulated so that the cell cycle was blocked in G_(0)/G_(1) phase.Conclusion The results demonstrated that CPX could inhibit human AGS gastric cancer cells growth and invasion,and which might be developed as a potential drug for gastric cancer treatment.