论文部分内容阅读
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 investigate the role of Ets5 (ETV5) in phenotypic transformation of vascular smooth muscle cells (VSMCs). Methods VSMCs were transfected with ETV5 overexpression adenovirus vector (Ad-ETV5 group) and green fluorescent protein (GFP) overexpression adenovirus vector (Ad-GFP group) respectively. Human ETV5-specific small interfering RNA (PDGF-BB + siRNAETV5 group) and its PDGF-BB + siRNAscramble group were transfected into VSMCs respectively and then PDGF-BB was used to induce VSMC phenotype transformation. Real-time quantitative PCR (qPCR) and Western blotting were used to detect the expression of α-smooth muscle actin (α-SMA) and myosin heavy chain 11 (MYH11) mRNA and protein of VSMC phenotype in each group. The migration and proliferation activities of VSMC in each group were detected by plate scratch test and CCK-8 kit, respectively. Results Compared with Ad-GFP group, the expression of α-SMA and MYH11 mRNA and protein in Ad-ETV5 group decreased (P <0.05) and the migration and proliferation activities increased (P <0.05). Compared with the PDGF-BB + siRNAscramble group, the expressions of α-SMA and MYH11 mRNA and protein in PDGF-BB + siRNAETV5 group were significantly increased (P <0.05), while the migration and proliferation were decreased (P <0.05). Conclusion ETV5 is involved in phenotypic transformation of VSMC.