MiR-26b enhances drug resistance of pancreatic cancer cells to gemcitabine by inhibiting p 53 gene expression
10.3969/j.issn.1673-4130.2017.22.001
- VernacularTitle:MiR-26b通过抑制p53基因表达增强胰腺癌细胞对吉西他滨抗药性
- Author:
Yanli WANG
1
;
Lei YANG
;
Yanmin CHANG
Author Information
1. 天津市中西医结合医院南开医院检验科
- Keywords:
pancreatic cancer;
p53;
gemcitabine;
drug resistance;
miR-26b
- From:
International Journal of Laboratory Medicine
2017;38(22):3073-3076
- CountryChina
- Language:Chinese
-
Abstract:
Objective To reveal the pancreatic cancer cell drug resistance mechanism to provide a basis for clinical treatment by studying miR-26b targeting p53 for promoting the drug resistance of pancreatic cancer cell line PANC-1 .Methods (1) The over-expression and knockdown plasmid of p53 and the fluorescent reporter vector containing 3′Untranslated region(3′UTR) were constructed respectively .(2) The effect of p53 and miR-26b on the growth and proliferation of PANC-1 was investigated by methyl thiazolyl tetrazolium(MTT) in the presence of gemcitabine .(3) The target relationship between miR-26b and p53 was determined by bioinformatics ,real-time polymerase chain reaction(PCR) ,fluorescent reporter vector and Western blot experiment .(4) The effect of p53 on the growth of miR-26b was investigated by salvage experiments .Results (1) The MTT experiment confirmed that ,in the presence of gemcitabine ,over-expression of p53 could inhibit the proliferation of pancreatic cancer cell line PANC-1 ,and knock-down of p53 could promote the growth and proliferation of PANC-1;(2) the bioinformatics prediction showed that miR-26b targeted p53 ,real-time PCR ,Western blot and fluorescent reporter vector experiment confirmed that p 53 is the target gene of miR-26b , and miR-26b inhibits transcription and translation of p53 by targeting the 3′UTR of p53 gene;(3) the MTT experiment confirmed that in the presence of gemcitabine ,over-expression of miR-26b could promote the growth and proliferation of PANC-1 cells ,and enhanced its resistance to gemcitabine ;(4) the rescue experiment confirmed that the simultaneous over-expression of p53 rescued the drug resistance promoting effect of miR-26b on PANC-1 .Conclusion miR-26b inhibits the expression of p53 by targeting 3′UTR of the p53 gene and enhances the drug resistance of pancreatic cancer cell line PANC-1 to gemcitabine .
