Screening of novel miRNAs targeting EZH2 3' untranslated region using lentivirus miRNAs library and their expressions in breast cancer cells and tissues.
- Author:
Cuicui LIU
1
;
Lulu WANG
;
Weiwei ZHAO
;
You PENG
;
Yuping WANG
;
Zhenliang SUN
;
Jing FENG
Author Information
- Publication Type:Journal Article
- MeSH: 3' Untranslated Regions; Breast Neoplasms; genetics; Cell Line, Tumor; Enhancer of Zeste Homolog 2 Protein; Female; Gene Expression Profiling; Humans; Lentivirus; genetics; MicroRNAs; genetics; Polycomb Repressive Complex 2; genetics
- From: Journal of Southern Medical University 2014;34(3):368-372
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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.
METHODSA 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.
RESULTSSeven 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).
CONCLUSIONNovel 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.
