紫草素对HGF诱导的人肺癌细胞上皮-间充质转化的逆转作用

Reverse effect of shikonin on epithelial-mesenchymal transition induced by HGF in lung cancer cells

目的:探讨紫草素(shikonin)对肝细胞生长因子(HGF)诱导的人非小细胞肺癌PC9细胞迁移、侵袭及上皮-间充质转化(EMT)的影响。方法:用HGF诱导PC9细胞建立EMT模型,采用不同剂量的shikonin干预24 h后,MTT法检测细胞活力;划痕愈合实验检测细胞的迁移能力;Transwell小室实验检测细胞的侵袭能力;Western blot法检测细胞中上皮型钙黏蛋白(E-cadherin)、神经型钙黏蛋白(N-cadherin)和波形蛋白(vimentin)的蛋白表达水平。结果:Shikonin可显著抑制PC9细胞的活力(P<0.01),随着给药剂量的增加,shikonin对细胞的生长抑制率显著上升,并呈一定的剂量依赖关系,IC_(50)为9.364μmol/L。HGF可诱导PC9细胞发生迁移和侵袭;划痕愈合实验和Transwell小室实验显示,shikonin能明显抑制由HGF诱导的肺癌PC9细胞迁移和侵袭(P<0.01)。Western blot检测结果显示HGF可诱导PC9细胞的EMT标志物E-cadherin蛋白表达下调,N-cadherin和vimentin蛋白表达上调,使其发生EMT;shikonin则可逆转由HGF诱导的PC9细胞E-cadherin蛋白表达下调及N-cadherin和vimentin蛋白表达上调(P<0.01)。结论:Shikonin能逆转由HGF诱导的肺癌PC9细胞EMT,同时抑制其迁移和侵袭。

AIM:To investigate the effects of shikonin on the migration,invasion and epithelial-mesenchymal transition(EMT)in human non-small-cell lung cancer PC9 cells induced by hepatocyte growth factor(HGF).ME-THODS:The effect of shikonin on the viability of PC9 cells was measured by MTT assay.The cell migration and invasion abilities were analyzed by wound healing assay and Transwell method,respectively.The protein expression levels of E-cadherin,N-cadherin and vimentin in the PC9 cells were determined by Western blot.RESULTS:The viability of PC9 cells was significantly inhibited by shikonin in a dose-dependent manner(P<0.01),with IC 50 at 9.364μmol/L.HGF significantly promoted the abilities of migration and invasion,and induced EMT in the PC9 cells.Shikonin significantly inhibited HGF-induced migration and invasion in the PC9 cells.The expression of E-cadherin was significantly down-regulated and the expression of N-cadherin and vimentin was significantly up-regulated in the presence of HGF(50μg/L).However,shikonin reversed HGF-induced EMT,as indicated by up-regulation of E-cadherin and down-regulation of N-cadherin and vimentin(P<0.01).CONCLUSION:Shikonin reverses HGF-induced EMT in lung cancer PC9 cells.