抑制HK-Ⅱ对人结肠癌细胞增殖和化疗敏感性的影响及机制

Effect and mechanism of HK-Ⅱ inhibition on cell proliferation and chemotherapy in human colon cancer cell lines

目的:探讨抑制HK-Ⅱ对人结肠癌细胞增殖和化疗敏感性的影响及可能机制。方法:应用免疫组织化学法检测HK-Ⅱ在人结肠癌细胞中的表达;采用MTT法观察HK-Ⅱ抑制剂(3-BrPA)对人结肠癌细胞增殖的影响和化疗增敏效应;通过Hoechst33258染色,用共聚焦激光扫描显微镜观察3-BrPA诱导的细胞凋亡;用流式细胞术、紫外分光光度计分别检测细胞死亡率、线粒体膜电位和线粒体膜通透转换孔(PTP)开放的变化。结果:HK-Ⅱ在人结肠癌细胞中阳性表达。3-BrPA能明显抑制人结肠癌细胞增殖,小剂量3-BrPA增加化疗敏感性。3-BrPA可诱导结肠癌细胞凋亡,引起线粒体膜电位下降和PTP开放。结论:抑制HK-Ⅱ可能是通过影响线粒体膜电位和PTP来发挥抗肿瘤作用。

OBJECTIVE： To study the role and mechanism of HK Ⅱ inhibition on cell proliferation and chemotherapy in human colon cancer ceil lines. METHODS： HK Ⅱexpression was evaluated by using theimmunohistoehemistry technique. The cell growth was assessed by MTT assay. Cell apoptosis was analysised hy u sing Hoeebst 33258 stain and confocal laser scanning microscopy. The cell death rate, mitochondria membrane vohage and opening of permeability transition pores （PTP） were explored by flow cytom etry and UV spectrophotometer analysis, respectively. RESULTS： HK Ⅱ expressions were observed in human colorectal cancer cell lines. HK Ⅱ specific inhibitor, 3-BrPA, signil＇icantly suppressed cellular growth, enhanced cytotoxic of chemotherapeutic drug and induced cellular apoptosis. The degression of mitochondria mem brane voltage and opening of PTP were also measured. CONCLU- SION： This study demonstrates that HK Ⅱinhibition may exert its anticancer activity via interfering with human colon cancer cell mitocbondria membrane voltage and opening of PTP.