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中国癌症防治杂志 ›› 2011, Vol. 3 ›› Issue (1): 5-9.doi: doi:10.3969/j.issn.1674-5671.2011.01.02

• 基础研究 • 上一篇    下一篇

野生型p53对SMMC-7721细胞中POLD1基因的影响

刘起理, 谭晓虹, 徐恒, 韦长元, 洪伟   

  • 出版日期:2011-03-25 发布日期:2011-04-25
  • 通讯作者: 徐恒、韦长元

The impact of p53 on POLD1 in SMMC-7721 cells

LIU Qi-Li, TAN Xiao-Hong, XU Heng, WEI Chang-Yuan, HONG Wei   

  • Online:2011-03-25 Published:2011-04-25
  • Supported by:

    国家自然科学基金资助项目(303296)

摘要: 目的探讨野生型p53基因表达对人肝癌细胞SMMC-7721中POLD1基因的影响。方法 设计并构建p53特异性小干扰shRNA绿色荧光真核表达质粒(p53-siRNA)和表达EGFPp53融 合 蛋 白的p53绿色荧光真核增强表达质粒(pEGFP-p53),通过稳定转染,将表达pEGFP-p53重组质粒、p53-siRNA转染入SMMC-7721细胞;经G418筛选,获得稳定细胞系7721-p53、7721-p53 RNAi。通过RT-PCR检测转染后p53、POLD1的mRNA。结果 在人肝癌细胞SMMC-7721中,野生型p53高表达组能够抑制POLD1的基因转录(P<0.001);而低表达组能够促进POLD1的基因转录(P<0.001)。结论 在人肝癌细胞SMMC-7721中,p53能够调控POLD1的基因转录。

关键词: SMMC-7721, POLD1, p53, RNA干扰

Abstract: ObjectiveTo investigate the impact of p53 on POLD1 gene in human liver cancer cell line SMMC-7721Methods Designed and constructed recombinant plasmid p53-siRNA and pEGFP-p53, then they  were transfected stably into SMMC-7721 cellsThrough G418 selection, the stable cell line 7721-p53,7721-p53RNAi  were acquiredThe p53 -RNA and POLD1 mRNA in these cell lines were detected by RT-PCR after transfectionResults High expression of wildtype p53 gene inhibited the transcription of POLD1 gene in SMMC-7721 cells, while low expression of p53 gene promoted the transcription of POLD1 geneConclusion p53 gene can regulate the transcription of POLD1 gene in SMMC-7721 cells

Key words: -SMMC-7721, POLD1, p53, RNA interference