| 摘要: |
| 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MYH11 mRNA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MYH11 mRNA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。 |
| 关键词: Ets差异基因5 血管平滑肌细胞 表型转化 细胞增殖 细胞迁移 |
| DOI：10.16781/j.0258-879x.2017.03.0312 |
| 投稿时间：2016-10-18修订日期：2017-02-15 |
| 基金项目: |
|
| Effect of transcription factor Ets variant gene 5 on phenotypic switching of vascular smooth muscle cells |
| SUN Xiang,ZHU Zhi-dong,ZENG Guang,CHEN Bing* |
(Cardiovascular Therapeutic Center, No. 117 Hospital of PLA, Hangzhou 310000, Zhejiang, China
*Corresponding author) |
| Abstract: |
| 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 ETV5-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 α-smooth muscle actin (α-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 α-SMA and MYH11 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. |
| Key words: Ets variant gene 5 vascular smooth muscle cells phenotypic switching cell proliferation cell migration |
