血红素加氧酶-1对肝癌细胞细胞周期调控因子的影响  

Effect of Heme Oxygenase-1 on Cell Cycle Regulatory Factors in Liver Cancer Cells

作　　者：高涵[1,2] 邹朝霞[1] 刘文杰[1] 张宏宇[1] 高旭[1] 

机构地区：[1]哈尔滨医科大学生物化学与分子生物学教研室,哈尔滨市150081 [2]齐齐哈尔医学院生物化学与分子生物学教研室,黑龙江省齐齐哈尔市161006

出　　处：《医学分子生物学杂志》2010年第2期99-103,共5页Journal of Medical Molecular Biology

基　　金：黑龙江省卫生厅基金（No.2009-211）,黑龙江省留学归国基金（No.LC06C21）,黑龙江省教育厅科技计划项目（No.115hz24）

摘　　要：目的观察血红素加氧酶-1（heme oxygenase 1，HO-1）对人肝癌细胞HepG2细胞周期调控因子的影响。方法构建含有野生型和突变型HO-1基因的重组载体pcDNA3．1（＋）-wtHO-1和pcDNA3．1（＋）-mHO-1G143H。利用脂质体介导的方法将构建好的重组载体转染肝癌细胞系HepG2，以空载体转染作为对照组。通过G418筛选建立稳定表达野生型和突变型HO-1的HepG2肝癌细胞系。经半定量RT—PCR、Western印迹检测转染细胞系中HO-1 mRNA和蛋白的表达水平。在HO-1表达改变的稳转细胞系中，利用Western印迹检测转染细胞系中P21、P27蛋白表达水平。结果成功实现了野生型和突变型HO-1在HepG2细胞中的过表达；野生型和突变型HO-1过表达均能诱导抑癌基因p21和p27的表达。结论HO．1过表达诱导抑癌基因p21和p27的表达与血红素分解产物无关。HO-1可能通过其它机制调节p21和p27的表达。Objective To elucidate the effect of heme oxygenase-1 on cell cycle regulator in HepG2 cell line. Methods To construct eukaryotic expression vectors expressing wild-type HO-1 and G143H mutant HO-1, named pcDNA3.1（ ＋ ） -wtHO-1 and pcDNA3.1（ ＋ ） - mHO-1G143H. HepG2 cells were transfected with wtHO-1 and mHO-1 using lipofectamine 2000. Stable transfected cells were selected by G418. Expression of HO-1 mRNA and protein were detected using RT-PCR and Western blot respectively. In addition, expression of p21 and p27, the major eyclin-dependent kinase （Cdk） inhibitors in the two cell lines were examined. Results HepG2 cell lines with wild type and mutant HO-1 overexpression were established successfully. Overexpression of both wild type and mutant HO-1 enhanced the expression of tumor suppressor P21 and P27. Conclusion HO- 1 overexpression upregulated the level of P21 and P27. This regulation is not dependent of hHO-1 catalytic activity, but probably other mechanisms.

关 键 词：血红素加氧酶-1 肝癌 细胞周期 细胞周期调控因子 

分 类 号：R349.53[医药卫生—基础医学]