医学分子生物学杂志2012，Vol.9Issue(2)：93-97,5.DOI:10.3870/j.issn.1672-8009.2012.02.004
p100蛋白表达抑制的 HepG2肝癌细胞稳定株的建立
Establishment of Human p100 Protein Stable Knockdown HepG2 Cell Line
摘要
Abstract
Objective To establish a p100 stable knockdown HepG2 cell line , and thus investigate the function of p 100 protein in hepatocarcinoma HepG2 cells. Methods The p100 shRNA expressing plasmid which carry eukaryotic selection marker Neo and GFP was transfected into HepG 2 cells using Lipofectamine. G418 resistant and GFP-positive single clones were picked up following term selection culture. Knockdown of p100 in the selected cell line was evaluated by western blot. Plate colony formation assay was used to detect the colony formation ability. MTS assay was used to analyze the cell proliferative response to cisplatin treatment. Cell migration was determined with wound healing assay. Results A p100 knockdown HepG2 stable cell line, HepG2 (p100I) , was successfully established , which provides an in vitro model to research the function of p 100 protein in hepatocellular carcinoma cells. Our experimental results showed that knockdown of p 100 protein in HepG2 cells reduced overall the colony formation , resistance to cisplatin induced cell death, and cell migration. Conclusion p100 protein affects several tumor related biological behaviors of HepG2 hepatocellular carcinoma cells , such as cell growth, cell survival and cell migration , thereby prob-ably contributing to liver cancer progression.关键词
人类p100蛋白/HepG2肝癌细胞/shRNA表达载体/G418耐药筛选/稳定株Key words
human p100 protein/ HepG2 cell line/ shRNA expressing plasmid / G418 selection: stable cell line分类
生物科学引用本文复制引用
田潇,尹洁,丁建民,史雪彬,何津岩,张毅,高星杰,经翔,杨洁..p100蛋白表达抑制的 HepG2肝癌细胞稳定株的建立[J].医学分子生物学杂志,2012,9(2):93-97,5.基金项目
国家高技术研究发展计划资助项目(863计划)(No.2007A,A02Z115),国家自然科基金(No.30970582,31100967,31170830),国家自然科学重大研究计划培育项目(No.90919032),国家教育部高等学校博士学科专项科研基金(No.20091202110001),中国博士后科学基金(No.2011M500529)天津市应用基 （863计划）
