摘要
目的研究乙肝病毒X蛋白结合蛋白(HBXIP)对卵巢癌细胞增殖功能的影响及其作用机制。方法将HBXIP质粒转染到卵巢癌细胞中,将细胞分为实验组(转染pCMV-HBXIP质粒)和对照组(转染pCMV-Tag2B质粒)。采用平板克隆形成实验和MTT实验检测卵巢癌细胞的增殖;荧光素酶报告基因实验检测转染HBXIP后Skp2启动子活性。采用PCR方法检测HBXIP在卵巢癌细胞中的表达;Western blotting检测转染HBXIP后卵巢癌细胞中HBXIP和Skp2蛋白的表达。结果平板克隆形成实验结果显示,实验组卵巢癌细胞克隆形成数高于对照组,差异有统计学意义(P<0.01)。MTT实验结果显示,实验组卵巢癌细胞的存活细胞数高于对照组,差异有统计学意义(P<0.05)。荧光素报告基因实验结果显示,与对照组相比,实验组中Skp2启动子活性显著升高(P<0.01)。PCR结果显示HBXIP基因在卵巢癌细胞中表达,但在对照组中表达欠缺。Western blotting结果显示,实验组卵巢癌细胞中HBXIP和Skp2的蛋白表达水平均高于对照组,差异有统计学意义(P<0.05)。结论 HBXIP可能通过调节Skp2的启动子活性和蛋白表达来促进卵巢癌细胞的增殖。
Objective To investigate the effect of hepatitis B X-interacting protein（HBXIP） on the proliferation of ovarian cancer cells and its underlying mechanism.Methods Ovarian cancer cells transfected with plasmid encoding pCMV-HBXIP were taken as experimental group,and those transfected with pCMV-Tag 2B vector served as control group.The proliferation of ovarian cancer cells was detected by colony formation assay and MTT assay.The Skp2 promoter activity in pCMV-HBXIP-transfected ovarian cancer cells was identified by luciferase report gene assay.The expression of HBXIP in ovarian cancer cells was detected by PCR.The expressions of HBXIP and Skp2 protein in ovarian cancer cells were assessed by Western blotting.Results Colony formation assay revealed that the formed colonies were significantly higher in the ovarian cancer cells of the experimental group than in control group（P0.01）.MTT assay indicated that the survival number was also significantly higher in the ovarian cancer cells of experimental group than in control group（P0.05）.Luciferase report gene assay showed that the Skp2 promoter activity was significantly increased in experimental group（P0.01）.The expression of HBXIP in ovarian cancer cells was detected with PCR,but none in control group.Western blotting revealed that the expression levels of HBXIP and Skp2 proteins were significantly increased in experimental group than in control group（P0.05）.Conclusion HBXIP may promote the proliferation of ovarian cancer cells by activating Skp2 promoter and up-regulating the expression of Skp2.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2013年第9期725-728,共4页
Medical Journal of Chinese People's Liberation Army
