Effect of Desmuslin gene over-expression on proliferation and migration of smooth muscle cells derived from primary varicose veins

Abstract

Objective To observe functional alterations in smooth muscle cells (SMCs) derived from varicose veins after over-expression of Desmuslin (DMN) gene by pRetroQ-AcGFP1-C1-DMN plasmid and further explore the effect of DMN gene involved in the development of varicose veins.Methods SMCs from varicose veins were cultured by the explant culture technique.The purity of the SMCs was identified by immunofluresence with a monoclonal antibody against human smooth muscle α-actin.DMN in SMCs was overexpressed by pRetroQ-AcGFP1-C1-DMN plasmid infected.The transfection efficiency was tested by real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting.CCK-8 assay was used to determine the proliferation of SMCs.Transwell assay was used to determine the migration of SMCs.Results Purity of these cultured SMCs was over 99％ identified by positive immunostaining with monoclonal SM-α-actin antibody.Levels of DMN mRNA and DMN protein in SMCs infected by pRetroQ-AcG-FP1-C1-DMN were significantly overexpressed compared with the empty vector group,(8.3 ± 1.1) × 104 and (4.5 ± 1.2) times respectively (P ＜ 0.01).The proliferation of SMCs of DMN over-expression group was (48.2 ± 4.5) ％ of the empty vector group by using CCK-8 assay (P ＜ 0.01).The migration ability of SMCs of DMN over-expression group was (44.3 ±4.7)％ of the empty vector group by using transwell assay (P ＜ 0.01).Conclusion The SMCs derived from varicose veins presented a significantly inhibited state of proliferation and migration after DMN over-expression,suggesting that DMN may play a key role in the development of varicose veins. Key words: Desmuslin; Retroviral vector; Vascular smooth muscle cells; Varicose veins