转录因子Ets差异基因5在血管平滑肌细胞表型转化中的作用

Effect of transcription factor Ets variant gene 5 on phenotypic switching of vascular smooth muscle cells

目的探讨转录因子Ets差异基因5(ETV5)在血管平滑肌细胞(VSMC)表型转化中的作用。方法分别采用ETV5过表达腺病毒载体(Ad-ETV5组)和绿色荧光蛋白(GFP)过表达腺病毒载体(Ad-GFP组)转染VSMC。人ETV5特异性小干扰RNA[血小板源性生长因子(PDGF)-BB+siRNAETV5组]及其随机阴性序列(PDGF-BB+siRNAscramble组)分别转染VSMC后采用PDGF-BB诱导VSMC表型转化。分别采用实时定量PCR(qPCR)和蛋白质印迹法检测各组细胞中VSMC表型标记物α-平滑肌肌动蛋白(α-SMA)和肌球蛋白重链11(MYH11)mRNA和蛋白的表达水平。分别采用平板划痕实验和CCK-8试剂盒检测各组VSMC的迁移和增殖活性。结果与Ad-GFP组相比,Ad-ETV5组VSMC中α-SMA和MYH11mRNA和蛋白的表达均降低(P<0.05),迁移和增殖活性均升高(P<0.05)。与PDGF-BB+siRNAscramble组相比,PDGF-BB+siRNAETV5组VSMC中α-SMA和MYH11mRNA和蛋白的表达均升高(P<0.05),而迁移和增殖活性降低(P<0.05)。结论 ETV5参与了VSMC的表型转化。

Objective To explore the effect of transcription factor E-twenty-six variant gene 5 （ETV5） on the phenotypic switching of vascular smooth muscle cells （VSMCs）. Methods The VSMCs were transfected with adenovirus vector overexpressing ETV5 in Ad-ETV5 group and green fluorescent protein （GFP） in Ad-GFP group. The VSMCs were transfected with human ETVS-specific small interfering RNA （siRNA） in platelet-derived growth factor （PDGF）-BB＋siRNAETV5 group and siRNA with random negative sequence in PDGF-BB＋ siRNAscramble group before inducing with PDGF-BB. The qPCR and Western blotting analysis were used to detect mRNA and protein levels of phenotypic switching markers of VSMC, including a-smooth muscle actin （a-SMA） and myosin heavy chain 11 （MYH11）. Scratch wound assay and CCK-8 assay were performed to identify the proliferation and migration of VSMCs. Results Compared with the Ad-GFP group, the expressions of mRNA and protein of a-SMA and MYHll were significantly decreased in the Ad-ETV5 group （P〈0. 05）, while the abilities of proliferation and migration were significantly improved （P〈0. 05）. Compared with PDGF-BB＋ siRNAscramble group, the expressions of mRNA and protein of α-SMA and MYH11 were significantly increased in the PDGF-BB＋siRNAETV5 group （P〈0. 05）, while the abilities of proliferation and migration were significantly inhibited （P〈0. 05）. Conclusion ETV5 may play an important role in the phenotypic switching of VSMCs.