[Targeting microRNA-mediated suppression of vascular endothelial growth factor gene expression and proliferation in malignant melanoma cells in vitro].

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.

• Authors: Yuan Jiang, Yongzhi Han, Jian Sun
• Journal: Nan fang yi ke da xue xue bao = Journal of Southern Medical University (Nan Fang Yi Ke Da Xue Xue Bao) Vol. 34 Issue 3 Pg. 358-63 (Mar 2014) ISSN: 1673-4254 [Print] China
• PMID: 24670449 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• MicroRNAs
• VEGFA protein, human
• Vascular Endothelial Growth Factor A

Topics

• Apoptosis
• Cell Line, Tumor
• Cell Proliferation
• Gene Expression Regulation, Neoplastic
• Genetic Vectors
• Humans
• Melanoma (genetics, metabolism, pathology)
• MicroRNAs (genetics)
• Skin Neoplasms
• Transfection
• Vascular Endothelial Growth Factor A (genetics, metabolism)
• Melanoma, Cutaneous Malignant