Screening of novel miRNAs targeting EZH2 3' untranslated region using lenti-miR virus library and their expressions in breast cancer cells and tissues
10.3969/j.issn.1673-4254.2014.03.16
- VernacularTitle:miRNAs慢病毒文库筛选靶向EZH23'非翻译区的新型miRNAs及其在乳腺癌中的表达
- Author:
Cuicui LIU
1
;
Lulu WANG
;
Weiwei ZHAO
;
You PENG
;
Yuping WANG
;
Zhenliang SUN
;
Jing FENG
Author Information
1. 南方医科大学第三临床医学院
- Keywords:
breast carcinoma;
EZH2;
3' untranslated region;
microRNA;
lenti-miR virus library
- From:
Journal of Southern Medical University
2014;(3):368-372
- CountryChina
- Language:Chinese
-
Abstract:
Objective To screen novel miRNAs targeting EZH2 3' untranslated region (UTR) in recombinational MCF-7 breast cancer cells over-expressing EZH2 3' UTR and quantitative analyze the expressions of the screened miRNA in breast cancer cells and tissues. Methods A lentiviral library was transfected into the recombinant cell line MCF-7. The cells were screened with cytotoxic agents before extraction of the genome for amplification of the miRNA precursors using PCR. The screened miRNAs were identified with sequence analysis and their expressions were analyzed quantitatively with real-time PCR in breast cancer cells and tissues. Results Seven miRNAs were screened from the recombinant MCF-7 cells, namely miR-15b, miR-16-2, miR-181b2, miR-217, miR-224, miR-329-1, and miR-487b, all of which failed to be predicted by bioinformatics software. Real-time PCR showed that miR-217, miR-329-1, and miR-487b were over-expressed in MCF-7 cells, and the expression of miR-15b and miR-16-2 was significantly increased in cancer tissues compared with the adjacent tissues (P<0.05). Conclusions Novel targeted miRNAs that can not be predicted by bioinformatics software were successfully screened from MCF-7 breast cancer cells over-expressing EZH2 3' UTR. These miRNAs are expressed differentially between normal breast cells and breast cancer tissues.
