黄独素B对结直肠癌细胞HCT116增殖及凋亡的影响

Effects of diosbulbin B on the proliferation and apoptosis of colorectal cancer cell HCT116

目的:探讨黄独素B(DB)对结直肠癌(CRC)细胞增殖及凋亡的影响,并研究其可能的作用机制。方法:首先将实验分为4组:对照组(0μmol/L)、DB低剂量组(10μmol/L)、DB中剂量组(20μmol/L)、DB高剂量组(40μmol/L),通过MTS法测定DB对CRC细胞HCT116活性的影响;通过划痕实验和单克隆实验,检测DB对HCT116细胞克隆形成、迁移和侵袭能力的影响;通过流式细胞术检测DB对细胞凋亡的影响;Western Blot检测DB对增殖、迁移、侵袭及凋亡等相关蛋白的表达。结果:与对照组比较,DB各剂量组能抑制HCT116细胞活性(P<0.05,P<0.01),且呈浓度依赖性;DB低、中、高剂量组能明显降低细胞克隆形成、迁移及侵袭能力,显著增加了细胞凋亡率;Western Blot显示,DB各剂量组能引起侵袭相关蛋白MMP-2、MMP-9的表达降低及凋亡相关蛋白Caspase-3、Caspase-9表达增加。结论:DB可能通过下调MMP-2、MMP-9蛋白和上调Caspase-3、Caspase-9凋亡蛋白表达来抑制HCT116细胞增殖、迁移和侵袭能力,诱导其凋亡,从而抑制CRC的发生发展。

Objective:To investigate the effect of diosbulbin B(DB)on the proliferation and apoptosis of colorectal cancer(CRC)cells,and to study its possible mechanism.Methods:Firstly,the experiment was divided into four groups:control group(0μmol/L),DB low-dose group(10μmol/L),DB medium-dose group(20μmol/L)and DB high-dose group(40μmol/L).The effect of DB on the activity of CRC HCT116 was determined by MTS method.The effects of DB on the clonal formation,migration and invasion of HCT116 cells were detected by scratch assay and monoclonal assay.The effect of DB on apoptosis was detected by flow cytometry.Western Blot was used to detect the expression of DB proteins related to proliferation,migration,invasion and apoptosis.Results:Compared with the control group,DB three dose groups could inhibit the activity of HCT116 cells(P<0.05,P<0.01),and there was a concentration-dependent manner.Meanwhile,cell clonal formation,migration,and invasion abilities were significantly reduced in the DB low,medium,high dose group.Western Blot results showed that DB three dose groups could cause the expression of MMP-2,MMP-9 related to proliferation and invasion to decrease,and the expression of apoptosis related protein Caspase-3,Caspase-9 to increase.Conclusion:DB may inhibit the proliferation,migration and invasion of HCT116 cells by down-regulating MMP-2,MMP-9 protein and up-regulating the expression of Caspase-3,Caspase-9 apoptotic protein,and induce its apoptosis,to inhibit the development of CRC.