miRNA-101 inhibits the expression of the enhancer of zeste homolog 2 in androgen-independent prostate cancer LNCaP cell line.
- Author:
Jian-xin LIU
;
Qi-fa ZHANG
;
Chang-hai TIAN
;
Yong ZHANG
;
Xiao-zhou HAN
;
Hao GUO
- Publication Type:Journal Article
- MeSH: Androgens; Cell Line, Tumor; Enhancer of Zeste Homolog 2 Protein; Genetic Vectors; Humans; Male; MicroRNAs; physiology; Polycomb Repressive Complex 2; genetics; metabolism; Prostatic Neoplasms; metabolism; RNA, Messenger; metabolism; Transfection
- From: National Journal of Andrology 2015;21(6):500-503
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the effect of miRNA-101 on the expression of the enhancer of zeste homolog 2 (EXH2) in human androgen-independent prostated cancer LNCaP cells.
METHODSWe divided LNCaP cells into a blank control, a negative control, and a miRNA-l01 transfection group, constructed the vector by transfecting synthetic miRNA-101 mimics into the LNCaP cells, and evaluated the efficiency of transfection by fluorescence microscopy. Then we determined the expression level of EZH2 mRNA by qRT-PCR in the three groups of cells and that of the EZH2 protein in the negative control and transfection groups by Western blot.
RESULTSGreen fluorescence signals were observed in over 70% of the LNCaP cells in the transfection group after 24 hours of transfection. At 72 hours, the expression of miRNA-101 was significantly upregulated in the transfected cells (P < 0.01), that of EZH2 mRNA was remarkably lower in the transfection group (0.01 ± 0.10) than in the blank control (0.95 ± 0.40) and negative control (0.86 ± 0.30) groups (both P < 0.01), and that of the EZH2 protein was increased in the negative control but decreased in the transfection group with the extension of culture time.
CONCLUSIONmiRNA-101, with its inhibitory effect on the expression of EZH2 in LNCaP cells, is a potential biotherapeutic for prostate cancer.