大葱油对人结肠癌SW480细胞增殖及转移能力的影响

Effects of Allium fistulosum L.oil on proliferation and metastasis of human colorectal cancer SW480 cells

目的:探讨大葱油对人结肠癌SW480细胞增殖和转移能力的影响。方法:采用MTT法、流式细胞仪检测不同浓度大葱油对SW480细胞生长能力及细胞周期的影响;采用Annexin V-FITC/PI双染检测细胞凋亡率;采用实时荧光定量PCR和Western blot法检测大葱油对细胞周期、凋亡和迁移侵袭相关基因和蛋白表达的影响。结果:大葱油能显著抑制SW480细胞生长,且呈剂量依赖性(P<0.01),100μg/ml大葱油作用SW480细胞24 h细胞抑制率显著高于人正常结肠CCD-18Co细胞(P<0.01);不同浓度的大葱油作用SW480细胞24 h,流式细胞仪检测结果显示细胞周期发生改变,细胞周期停滞于G2/M期,其G2/M期细胞比率分别从10.43%增至15.69%(50μg/ml)和24.07%(100μg/ml)且诱导细胞发生凋亡。25、50和100μg/ml大葱油作用SW480细胞24 h后,实时荧光定量PCR和Western blot检测与细胞周期、凋亡、迁移侵袭相关蛋白Cyclin D1、CDK4、CDK6、Bcl-2、MMP2、MMP9、LIMK1p-Thr508和Cofilinp-Ser3基因和蛋白表达量呈下降趋势,Caspase-9、Caspase-3和Bax基因和蛋白表达量呈上升趋势(P<0.05)。结论:大葱油具有抑制人结肠癌SW480细胞生长及转移能力,诱导G2/M期阻滞,并促进其凋亡。

Objective:To investigate the effect and molecular mechanism of Allium fistulosum L.oil on proliferation and metastasis of human colorectal cancer SW480 cells.Methods:SW480 cells were incubated with different concentrations(25,50,100μg/ml)of Allium fistulosum L.oil for 48 h and then the capability of cellular proliferation were detected by MTT assay.Meanwhile,the cell-cycle analysis and cell apoptosis were determined by flow cytometry.Real-time quantitative PCR and Western blot were applied to detect the expression level of mR NAs and proteins related to cell cycle,cell apoptosis and metastasis,respectively.Results:Allium fistulosum L.oil significantly inhibited proliferation of SW480 cells in a concentration dependent manner than that in CCD-18 Co cells.The cell viability of SW480 cells exposed to 100μg/ml Allium fistulosum L.oil was significantly lower than that in control group(P<0.01).Allium fistulosum L.oil prominently induced a dose-dependent increase in the percentage of cells in the G2/M phase compared with the control.The percentage of G2/M phase cells was 10.43%,15.69%and 24.07%when SW480 cells were treated with DMSO,50μg/ml and 100μg/ml of Allium fistulosum L.oil,respectively.Allium fistulosum L.oil-induced SW480 cell apoptosis was significantly increased than that of the control cells.After exposure to 25,50,100μg/ml Allium fistulosum L.oil,the mR NA and protein expression levels of Cyclin D1,CDK4,CDK6,Bcl-2,MMP2,MMP9,LIMK1 p-Thr508 and Cofilinp-Ser3 significantly decreased than those in control group,while the mR NA and protein expression levels of Caspase-9,Caspase-3 and Bax significantly increased than those in control group(P<0.05).Conclusions:Allium fistulosum L.oil can inhibit the proliferation of human colorectal cancer SW480 cells by arresting SW480 cells at the G2/M phase and promoting cell apoptosis.And it also can inhibite the metastasis of SW480 cells by regulating the expression of proteins related to metastasis.