忍冬藤提取物抗结肠癌作用研究

Effect of Lonicera japonica extract on colon cancer

目的研究忍冬藤提取物的体外抗结肠癌作用。方法体外培养结肠癌HCT116和SW480细胞并用忍冬藤提取物干预,采用CCK8法检测忍冬藤提取物对HCT116和SW480细胞增殖的影响;采用Hochest法和Annexin V/PI法检测忍冬藤提取物及p53抑制剂PFT-α对HCT116和SW480细胞凋亡的影响;JC-1法检测忍冬藤提取物对HCT116细胞线粒体膜电位的影响;Western Blot检测忍冬藤提取物对HCT116细胞相关蛋白表达水平的影响。结果相较于空白组,忍冬藤提取物可明显抑制结肠癌HCT116和SW480细胞增殖,并促进细胞凋亡(P<0.01),促进HCT116细胞线粒体膜电位坍塌,明显上调HCT116细胞Bax/Bcl-2的比值(P<0.01)以及Cleaved PARP、Cyt-C、Cleaved Caspase-3和p53蛋白表达(P<0.05,P<0.01);相较于忍冬藤提取物组,忍冬藤提取物+PFT-α明显抑制HCT116细胞凋亡(P<0.01)。结论忍冬藤提取物可通过诱导p53依赖的线粒体凋亡发挥抗结肠癌作用。

Objective To investigate the effect of Lonicera japonica extract on colon cancer.Methods Colon cancer HCT116 and SW480 cells were cultured in vitro and treated with L.japonica extract.CCK8 assay was used to detect the effect of L.japonica extract on the proliferation of HCT116 and SW480 cells.Hochest assay and Annexin V/PI assay were used to detect the effect of methanol extract of L.japonica and PFT-αon apoptosis of HCT116 and SW480 cells.JC-1 method was used to detect the effect of L.japonica extract on mitochondrial membrane potential(MMP)of HCT116 cells.Western blot was used to detect the effect of L.japonica extract on the expression level of related proteins in HCT116 cells.Results Compared with the control group,extract of L.japonica inhibited the proliferation of colon cancer HCT116 and SW480 cells,induced the apoptosis of SW480 and HCT116 cells significantly(P<0.01).The extract of L.japonica changed the MMP in HCT116 cells,promoted the ratio of Bax/Bcl-2 and the protein expression of Cleaved PARP,Cyt-C,Cleaved Caspase-3,p53(P<0.05,P<0.01)in HCT116 cells.Compared with the extract of L.japonica group,extract of L.japonica and PFT-αinhibited the apoptosis of HCT116 cells obviously.Conclusions Extract of L.japonica could induce the apoptosis of p53-dependent mitochondria,which plays an anti-colon cancer role.