Down-regulation of miR-21 expression enhances the radiosensitivity of TE-1 cells in vitro.
- Author:
Xiaoqing LI
1
;
Xin CHEN
;
Shan HUANG
;
Shaomin CHE
;
Xiaozhi ZHANG
Author Information
- Publication Type:Journal Article
- MeSH: Cell Line, Tumor; Down-Regulation; Genetic Vectors; Humans; Lentivirus; genetics; MicroRNAs; genetics; metabolism; Radiation Tolerance; Wnt Proteins; metabolism; Wnt Signaling Pathway; beta Catenin; metabolism
- From: Journal of Southern Medical University 2012;32(11):1559-1563
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the effect of miR-21 down-regulation on the radiosensitivity of TE-1 cells in vitro.
METHODSTE-1 cells were transfected via lentivirus with a vector containing the antisense oligonucleotides of miR21, and the subclones with stable down-regulation of miR21 expression were selected with puromycin and designated as TE-1-miR21(-), whose expression level of miR21 was determined using real-time quantitative PCR. The radiosensitivity of TE-1 and TE-1-miR21(-) cells were evaluated with colony formation assay, and the expressions of β-catenin was determined using Western blotting and RT-PCR. Flow cytometry was used to analyze the proportion of p75NTR(+) cells in TE-1 and TE-1-miR21(-) cells.
RESULTSA cell subclone stably expressing a low level of miR21 was obtained and verified by real-time quantitative PCR. Colony formation assay showed an enhanced the radiosensitivity of TE-1-miR21(-) cells compared to parental TE-1 cells. RT-PCR revealed no significant changes in β-catenin mRNA expression in TE-1-miR21(-) cells, whereas its β-catenin protein expression was markedly suppressed by high-dose (8 and 10 Gy) irradiation. Flow cytometry assay showed a decreased proportion of p75NTR(+) cells in TE-1-miR21(-) cells compared to that in TE-1 cells.
CONCLUSIONDown-regulation of miR21 can enhance the radiosensitivity of TE-1 cells, which might result from the inactivation of wnt/β-catenin signal pathway and a decreased p75NTR(+) cell proportion.
