Targeting microRNA-mediated suppression of vascular endothelial growth factor gene expression and proliferation in malignant melanoma cells in vitro
10.3969/j.issn.1673-4254.2014.03.14
- VernacularTitle:靶向VEGF的miRNA对恶性黑色素细胞增殖和凋亡的影响
- Author:
Yuan JIANG
1
;
Yongzhi HAN
;
Jian SUN
Author Information
1. 广东省人民医院//广东省医学科学院皮肤科
- Keywords:
malignant melanoma;
miRNA;
vascular endothelial growth factor;
cell proliferation;
apoptosis
- From:
Journal of Southern Medical University
2014;(3):358-363
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the inhibitory effect of targeting miRNA on the expression of vascular endothelial growth factor (VEGF) and cell proliferation in malignant melanoma (MM) SKmel-28 cells. Methods Recombination miRNA plasmid vectors targeting VEGF gene were transfected into SKmel-28 cells via Lipofectamine 2000. The integrity of the inserted fragments was detected using colony PCR and sequence analysis. The expression of VEGF mRNA and protein in SKmel-28 cells was detected by RT-PCR and Western blotting, respectively. MTS assay was used to determine the inhibitory effect of a selected targeting miRNA on SKmel-28 cell proliferation, and the apoptosis of SKmel-28 cells was detected using flow cytometry. Results Transfection with the targeting miRNAs significantly down-regulated the expressions of VEGF mRNA and protein in SKmel-28 cells (P<0.01), and the miRNA construct X-26-2n-1 showed the highest inhibitory effect. The miRNA X-26-2n-1 significantly suppressed SKmel-28 cell proliferation in a time-dependent manner (P<0.01) and increased the early, late and overall apoptosis rates of the cells (P<0.01). Conclusion The targeting miRNA we constructed can effectively suppress the cell proliferation and induce apoptosis of SKmel-28 cells by down-regulating the expressions of VEGF gene.
