Effect of p53 with R213 mutationon the expression of p21 in colorectal cancer cells HCT116
- VernacularTitle:结肠癌细胞HCT116中p53的R213突变对靶基因p21表达的影响
- Author:
Yan ZHANG
;
Yanjun ZHANG
;
Ye ZHANG
- Keywords:
p53;
arginine213;
p21;
mutation
- From:
Basic & Clinical Medicine
2017;37(5):663-667
- CountryChina
- Language:Chinese
-
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.
