广藿香酮对肺癌PC-9细胞增殖和凋亡的影响

Effect of pogostone on the proliferation and apoptosis of lung cancer PC-9 cells

目的:探讨广藿香酮对肺癌PC-9细胞增殖、克隆形成、侵袭、迁移和凋亡的影响,并初步探讨其可能的分子机制。方法:配制不同浓度的广藿香酮(50,100,150,200,250,300,350,400μmol·L^(-1))处理人肺癌PC-9、A549和95D细胞24,48 h,采用CCK-8法检测细胞增殖活力,筛选出其中对广藿香酮最敏感的细胞株并确定低、中、高浓度用于后续实验;将PC-9细胞分为对照组和广藿香酮低、中、高浓度组,采用平板克隆实验、Transwell小室实验、划痕愈合实验和流式细胞术分别检测细胞克隆形成、侵袭、迁移能力和细胞凋亡;采用Western blot检测PC-9细胞中PCNA、Vimentin、MMP-2、MMP-9、Bax、Bcl-2、Caspase-3、Cleaved-caspase-3蛋白的表达。结果:广藿香酮可有效抑制PC-9、A549和95D细胞增殖能力,且呈剂量和时间依赖性,其中PC-9细胞对广藿香酮最敏感;广藿香酮浓度分别为100,150,200μmol·L^(-1)处理PC-9细胞48 h,与对照组比较,广藿香酮低、中、高浓度组中细胞克隆形成、侵袭和迁移能力显著下降,细胞凋亡率显著上升;Western blot结果显示,与对照组比较,广藿香酮低、中、高浓度组PC-9细胞中PCNA、Vimentin、MMP-2、MMP-9和Bcl-2蛋白的表达显著减少,Bax、Cleaved-caspase-3蛋白的表达显著增加。结论:广藿香酮能显著抑制肺癌PC-9细胞的增殖、克隆形成、侵袭和迁移能力,并诱导细胞凋亡,分子机制可能与下调细胞中PCNA、Vimentin、MMP-2、MMP-9和Bcl-2蛋白的表达,上调Bax、Cleaved-caspase-3蛋白的表达有关。

OBJECTIVE To investigate the effects of pogostone on the proliferation,clonogenicity,invasion,migration and apoptosis of lung cancer PC-9 cells,and to preliminarily explore the possible molecular mechanism.METHODS Different concentrations of pogostone(50,100,150,200,250,300,350,400μmol·L^(-1))were used to treat human lung cancer PC-9,A549 and 95D cells for 24 and 48 h.The cell proliferation activity was detected by CCK-8 method,the cell lines most sensitive to pogostone were selected,and low,medium and high concentrations were determined for subsequent experiments;PC-9 cells were divided into control group and pogostone low,medium and high concentration groups,and clonogenic assay,Transwell chamber assay,scratch healing assay and flow cytometry were used to respectively determine clonogenic potential,invasion,migration ability and apoptosis of PC-9 cells.The protein levels of PCNA,Vimentin,MMP-2,MMP-9,Bax,Bcl-2,Caspase-3 and Cleaved-caspase-3 in PC-9 cells were determined by Western blot.RESULTS Pogostone inhibited the proliferation of PC-9,A549 and 95D cells in a dose-dependent and time-dependent manner,and PC-9 cells were the most sensitive to pogostone.The PC-9 cells were treated with 100,150,and 200μmol·L^(-1) pogostone for 48 h,and compared with those in the control group,the cell clone formation,invasion and migration ability were significantly reduced and the apoptosis rate was significantly increased in pogostone low,medium and high concentration groups.The Western blot results showed that the expression of PCNA,Vimentin,MMP-2,MMP-9 and Bcl-2 proteins were significantly reduced and the expression of Bax and Cleaved-caspase-3 protein was significantly increased in PC-9 cells in pogostone low,medium and high concentration groups compared to those in the control group.CONCLUSION Pogostone can significantly inhibit the proliferation,colony formation,invasion and migration of PC-9 cells,and induce the cell apoptosis.The molecular mechanism may be related to the down-regulation of PCNA,Vimentin,MMP-2,MMP-9 and Bcl-2 protein expression,and up-regulation of Bax and Cleaved-caspase-3 protein expression.