MiR-21 regulates the invasion of U251 human glioblastoma cells in vitro
10.3760/cma.j.issn.1671-8925.2010.10.007
- VernacularTitle:miR-21调控人脑胶质瘤细胞侵袭能力的体外研究
- Author:
Xuan ZHOU
1
;
Yu REN
;
Guang-Xiu WANG
;
Pei-Yu PU
;
Chun-Sheng KANG
Author Information
1. 天津医科大学肿瘤医院
- Keywords:
RNA interference;
Invasion;
U251 glioblastoma cell line;
Gene therapy
- From:
Chinese Journal of Neuromedicine
2010;09(10):991-995
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the mechanism of miR-21 in regulating the invasion of human glioblastoma (GBM) cells in vitro. Methods The transfection reagent oligofectamine was mixed with antisense miRNA-21 (AS-miR-21) and nonsense oligodeoxyribonucleotides (ODN), respectively, and then, they were added into the medium of U251 GBM cell line as AS-miR-21 treatment group and nonsense ODN treatment group, respectively; control group (treated with PBS) was also established.MiR-21 luciferase reporter assay was used to detect the miR-21 knocking down effect. Matrigel cell growth assay and Transwell assay were used to determine the invasion and migration abilities of U251 cells. Western blotting was employed to test the expressions of invasion-related proteins (FAK,MMP-9/2, TIMP-1 and Tubulin-α); immunofluorescence was also employed to observe the morphology of Tubulin-α protein in GBM cells. Results Luciferase intensity in as-miR-21 treated U251 cells was significantly suppressed as compared with that in the control group and nonsense ODN treatment group (P＜0.05). The diameter of cultured clone in as-miR-21 treated U251 cells was smaller than that in the controls and nonsense ODN treatment group (F=102.819, P=0.000). Decreased cells via the transwell member in thc AS-miR-21 treatment group were detected as compared with those in the controls and cnonsensc ODN treatment group (F=243.465, P=0.000). The expressions of FAK, MMP-2/9 weredown-regulated and that of TIMP-1 was up-regulated in the AS-miR-21 treated tumor cells as compared with the other 2 groups (P＜0.05). No obvious changes were noted on the expression of Tubulin α,however, the morphology of Tubulin α protein in the AS-miR-21 treatment group changed. Conclusion High expression of miR-21 induce the ability of U251 GBM cell invasion and miR-21 can be taken as a candidate for gene therapy of human glioma.
