紫草素对人结肠癌细胞HCT116自噬和凋亡的影响

目的:研究紫草素对人结肠癌细胞HCT116自噬和凋亡的影响。方法:以0(空白对照)、10、20、40μg/mL紫草素作用于HCT116细胞48 h后,采用MTT法检测细胞增殖抑制率;采用流式细胞术检测细胞凋亡率;分别采用实时荧光定量-聚合酶链式反应法和Western blotting法检测细胞中微管相关蛋白轻链3(LC3)和自噬相关蛋白Beclin-1、p62的mRNA及其蛋白表达水平。结果:与空白对照比较,10、20、40μg/mL紫草素作用48 h后HCT116细胞的增殖抑制率和凋亡率均显著升高(P<0.05或P<0.01)。10、20、40μg/mL紫草素作用于HCT116细胞48 h后均可不同程度地升高细胞中LC3、Beclin-1、p62 mRNA及其蛋白的表达水平,除10μg/mL紫草素作用后细胞中LC3 m RNA表达水平升高不显著外,其余指标水平与空白对照比较差异均有统计学意义(P<0.05或P<0.01)。结论:紫草素可诱导人结肠癌细胞HCT116发生凋亡,并激活其自噬途径。

Effects of Shikonin on Autophagy and Apoptosis of Human Colon Cancer HCT116 Cells

OBJECTIVE:To study the effects of shikonin on autophagy and apoptosis of human colon cancer HCT116 cells.METHODS:After treating HCT116 cells for 48 h with shikonin at 0(blank control)10,20,40μg/mL,MTT method was used to detect inhibitory rate of cell proliferation.Flow cytometry was used to detect cell apoptosis rate.RT-qPCR assay and Western blotting were respectively used to detect m RNA and protein expressions of microtubule associated protein light chain 3(LC3)and autophagy-related protein Beclin-1 and p62.RESULTS:Compared with blank control,after treated with 10,20,40μg/mL shikonin for 48 h,proliferation inhibitory rate and apoptosis rate of HCT116 cells were increased significantly(P<0.05 or P<0.01).After treated with 10,20,40μg/mL shikonin for 48 h,mRNA and protein expressions of LC3,Beclin-1 and p62 in HCT116 cells were increased to different extents;except that mRNA expression of LC3 was not increased significantly after treated with 10μg/mL shikonin,the difference were statistically significant in other indexes,compared with blank control(P<0.05 or P<0.01).CONCLUSIONS:Shikonin can induce the apoptosis of human colon cancer HCT116 cells and activate its autophagy pathway.