槲皮素对HSP-27敲减结肠癌SW480细胞增殖、迁移和侵袭的影响及作用机制研究

Effect of Quercetin on Proliferation, Migration and Invasion in Colon Cancer SW480 Cells with HSP-27 Knockdown and Its Mechanism

目的:研究槲皮素对HSP-27敲减结肠癌SW480细胞增殖、迁移和侵袭的影响,并探讨其作用机制。方法:通过脂质体Lipofectamine 2000将HSP-27-siRNA导入SW480细胞,采用RT-qPCR测定转染细胞中HSP-27基因表达。将SW480细胞分为空白对照组、阴性siRNA转染组(NC组)、HSP-27-siRNA转染组(siRNA组)、阴性siRNA转染+槲皮素组(NC+槲皮素组)、HSP-27-siRNA转染+槲皮素组(siRNA+槲皮素组),分别采用CCK-8法、Transwell实验和划痕实验检测槲皮素对HSP-27敲减SW480细胞增殖、侵袭、迁移和凋亡的影响,并采用RT-qPCR和Western blotting法测定槲皮素对HSP-27敲减SW480细胞Bcl-2、Bax mRNA和蛋白的影响。结果:SW480细胞经HSP-27 siRNA转染后HSP-27 mRNA表达显著降低(P<0.05);与NC组比较,siRNA组及NC+槲皮素组SW480细胞增殖抑制率显著升高,细胞侵袭数显著减少,细胞划痕愈合率显著降低,凋亡率显著升高,Bcl-2 mRNA及蛋白表达显著降低,Bax mRNA及蛋白表达显著升高(P<0.05)。与NC+槲皮素组比较,siRNA+槲皮素组SW480细胞增殖抑制率显著升高,细胞侵袭数显著减少,细胞划痕愈合率显著降低,凋亡率显著升高,Bcl-2 mRNA及蛋白表达显著降低,Bax mRNA及蛋白表达显著升高(P<0.05)。结论:槲皮素对HSP-27敲减SW480细胞增殖、侵袭和迁移具有抑制作用,其机制可能与增加HSP-27敲减SW480细胞的凋亡,并调节Bcl-2、Bax mRNA及蛋白的表达有关。

Objective:To explore the effect of quercetin on proliferation,migration and invasion in colon cancer SW480 cells with HSP-27 knockdown and to investigate its mechanism.Methods:Lipofectamine 2000 was used to transfer HSP-27-siRNA into SW480 cells,and the expression of HSP-27 gene in transfected cells was determined by RT-qPCR.SW480 cells were divided into normal control group,negative siRNA transfection group(NC group),HSP-27-siRNA transfection group(siRNA group),quercetin combined with negative siRNA transfection group(NC+quercetin group),quercetin combined with HSP-27-siRNA transfection group(siRNA+quercetin group).The effects of quercetin on proliferation,invasion,migration and apoptosis of SW480 cells with HSP-27 knockdown were detected by CCK-8 method,Transwell experiment,scratch experiment,respectively.The effects of quercetin on Bcl-2 and Bax mRNA and proteins in SW480 cells with HSP-27 knockdown were determined by RT-qPCR and Western blotting methods.Results:HSP-27 mRNA expression in SW480 cells was significantly decreased after transfection with HSP-27 siRNA(P<0.05).Compared with NC group,the proliferation inhibition rate of SW480 cells in siRNA group and NC+quercetin group was significantly induced,the number of cell invasion was significantly decreased,the rate of cell scratch healing was significantly decreased,the rate of apoptosis was significantly increased.Bcl-2 mRNA and protein expressions were significantly decreased,while Bax mRNA and protein expressions were significantly increased(P<0.05).Compared with NC+quercetin group,the proliferation inhibiton rate of SW480 cells in siRNA+quercetin group was significantly increased,the number of cell invasion was significantly decreased,the rate of cell scratch healing was significantly decreased,the rate of apoptosis was significantly increased,the expressions of Bcl-2 mRNA and protein were significantly decreased,and the expressions of Bax mRNA and protein were significantly increased(P<0.05).Conclusion:Quercetin can inhibit proliferation,invasion and migration of SW480 cells with HSP-27 knockdown,its mechanism may be related to the increase of apoptosis in SW480 cells with HSP-27 knockdown,and the regulation of Bcl-2,Bax mRNA and protein expressions.