Introduction of plasmid-mediated exogenous microRNA to silence PTTG1 gene expression and inhibit proliferation and invasiveness of glioma cells HUANG Qing-feng, LU Yi-eheng, BAI
10.3760/cma.j.issn.1671-8925.2008.08.001
- VernacularTitle:质粒介导外源性微小RNA干扰PTTG1表达抑制恶性人脑胶质瘤细胞增殖和侵袭
- Author:
Qing-Feng HUANG
1
;
Yi-Cheng LU
;
Ru-Lin BAI
;
Chun LUO
;
Guo-Han HU
Author Information
1. 第二军医大学附属长征医院
- Keywords:
PTTG1;
MicroRNA;
Glioma;
Cell proliferation;
Tumor infiltration
- From:
Chinese Journal of Neuromedicine
2008;7(8):757-761
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the role of pituitary tumor transforming gene 1 (PTTG1) in the growth and invasion of human glioma cell line by introduction of exogenous microRNA to silence PTTG 1 gene expression. Methods Two double-stranded DNA pcDNA6.2-GW/EmGFP-miR vectors (MIR-1, MIR-2) targeting human PTTG1 mRNA and a negative control plasmid (Neg) were constructed, and were transfected into human U251 cells with high metastatic potentials. Real-time PCR and Western blotting were used to quantify the mRNA and protein levels of PTTG1, respectively. Proliferation and invasiveness of transfected U251 cells were analyzed by MTT assay and Matrigel invasion assay. Results After transfection, Expression of PTTG1 mR.NA was inbibited significantly with inhibitory rates of 87.6% in MIR-2 group, and the protein levels were significantly lower than those of the other groups. There was significant difference in cellular growth rate among the 3 groups. The growth inhibiting rates in the MIR-2 group are 10.7%-34.7%. The migrating number of U251 cells transfected with MIR-2 with relative percentage (12.3±1.0)% was also significantly decreased as compared the Neg group (24.7±1.4)% and Mock group (24.0±2.0)%. Conclusion Introduction of exogenous miRNA to U251 cell line by transfection of MIR-2 can effectively reduce the PTTG1 expression, which can significantly inhibit the proliferation and decrease the invasiveness of glioma cells.
