中国免疫学杂志2011，Vol.27Issue(2)：120-125,6.DOI:10.3969/j.issn.1000-484X.2011.02.006
过表达HSPC238对Bel7402细胞周期的影响
Effect of HSPC238 overexpression on cell cycle of human hepatoma carcinoma cell line Bel7402
摘要
Abstract
Objective:To construct the recombinant eukaryotic expressing vector haboring human gene HSPC238,and to study the effect of HSPC238 overexpression on the cell cycle of Bel7402. Methods: The cDNA of HSPC238 was amplified from the recombinant plasmid pET28b/HSPC238 by PCR method. After sequencing, the target gene frame of HSPC238 cDNA was subcloned into eukaryotic expression vector pcDNA3.1 (-). The recombinant plasmid pcDNA3.1 (-)/HSPC238 was transfected into Bel7402 cell line by lipofectamine 2000. Positive clones were screened by G418 for 4 weeks. Then,the mRNA and protein expression levels of HSPC238 in positive clones were detected by RT-PCR and Western blot,respectively. Finally,cell cycle was analyzed by flow cytometry after synchronization of cell cycle by starvation method. Results:The construction of the recombinant plasmid pcDNA3.1(-)/HSPC238 was proved by restriction enzyme digestion analysis and DNA sequencing. Compared with pcDNA3.1(-) transfected group and untransfected group, the expression of HSPC238 in pcDNA3.1 (-)/HSPC238 transfected group was significantly increased both in mRNA level and in protein level. Flow cytometry suggested that HSPC238 could postpone the velocity of HSPC238-transfected Be17402 cell from phase G1 into phase S.Conclusion:The eukaryotic expression plasmid pcDNA3.1(-)/HSPC238 has been successfully constructed and HSPC238 is overexpressed in Bel7402 cell line. Our research suggests that overexpression of HSPC238 could postpone the cell cycle of Be17402. This will benefit further studies on the function of human gene HSPC238.关键词
HSPC238/过表达/Bel7402/细胞周期分类
医药卫生引用本文复制引用
黄湘,谭家余,王穗海,李明..过表达HSPC238对Bel7402细胞周期的影响[J].中国免疫学杂志,2011,27(2):120-125,6.基金项目
本文为国家高技术研究发展计划(863)资助项目(2006AA02A311)和广东省科技计划项目(2010B031600307) （863）
