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结肠癌细胞HCT116中p53的R213突变对靶基因p21表达的影响

张妍 张艳君 张业

基础医学与临床2017,Vol.37Issue(5):663-667,5.
基础医学与临床2017,Vol.37Issue(5):663-667,5.

结肠癌细胞HCT116中p53的R213突变对靶基因p21表达的影响

Effect of p53 with R213 mutationon the expression of p21 in colorectal cancer cells HCT116

张妍 1张艳君 2张业3

作者信息

  • 1. 首都医科大学附属北京潞河医院 中心实验室, 北京 101149
  • 2. 首都医科大学附属北京潞河医院 消化内科, 北京 101149
  • 3. 中国医学科学院北京协和医学院 基础医学研究所 医学分子生物学国家重点实验室, 北京 100005
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摘要

Abstract

Objective To determine impacts of p53 with R213 mutation on the expression of its downstream target gene p21 in HCT116-/-colorectal cancer cell line.Methods Mutations of p53 at R213Q (R/Q) and R213K (R/K) were respectively constructed and transfected into the p53 null HCT116 cells.The promoter activity drived by p53 was detected by dual-luciferase reporter system.EMSA assay was used to detect the ability of p53 binding to the promoter of its target gene p21.The expression of p53 downstream target genes p21 was examined by real-time PCR and Western blot experiments.Results The mutation of R213K disrupted the efficiency of p53 transactivation(P<0.05).The mutants were less efficient in targeting the consensus binding sequences of p53 in the regulatory region of p21 gene(P<0.05), In addition, The R/K mutant attenuated the expression of p21 gene(P<0.05).The mobility of R213Q was increased.Conclusions The different mutations of Arginine 213 have different impact on p21 in colorectal cancer cells.It could exist diverse regulating mechanisms.

关键词

p53/213位精氨酸/p21/突变

Key words

p53/arginine213/p21/mutation

分类

生物科学

引用本文复制引用

张妍,张艳君,张业..结肠癌细胞HCT116中p53的R213突变对靶基因p21表达的影响[J].基础医学与临床,2017,37(5):663-667,5.

基金项目

国家自然科学基金(31671359) (31671359)

基础医学与临床

OACSCDCSTPCD

1001-6325

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