The role of Exo70 in vascular smooth muscle cell migration.
The role of Exo70 in vascular smooth muscle cell migration.
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DOI：
10.1186/s11658-016-0019-8
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发表时间：
2016
影响因子：
8.3
通讯作者：
Yin M
中科院分区：
文献类型：
--
作者：
Ma W;Wang Y;Yao X;Xu Z;An L;Yin M
Exo 70是外囊复合物的关键亚基，具有高度保守的序列，广泛存在于酵母、哺乳动物和植物中。在酵母中，Exo 70介导胞吐过程并促进囊泡与质膜的锚定和整合。在哺乳动物细胞中，Exo 70参与维持细胞形态、细胞迁移、细胞连接、mRNA剪接和其他生理过程，以及参与胞吐作用。然而，Exo 70在哺乳动物细胞中的功能尚未得到充分认识。本文研究了大鼠血管平滑肌细胞系A7 r5中Exo 70的表达及其在细胞迁移中的作用。免疫荧光分析显示Exo 70、α-actin和tubulin在A7 r5细胞中的表达存在共定位，我们用细胞松弛素B处理A7 r5细胞以去除α-actin，以进一步证实Exo 70和α-actin的共定位。我们通过伤口愈合试验分析了Exo 70与肌动蛋白在迁移细胞边缘的共定位，以确定Exo 70是否可能在细胞迁移中发挥作用。接下来，我们通过伤口愈合实验和transwell实验分析了RNAi沉默前后A7 r5细胞的迁移和侵袭能力。Exo 70与细胞骨架相互作用的机制可以通过免疫沉淀技术和创伤愈合实验来阐明。结果表明，Exo 70和α-actin共定位于迁移细胞的前缘。当通过RNAi沉默Exo 70表达时，A7 r5进行细胞迁移的能力降低。与正常细胞相比，减少RNAi处理的A7 r5细胞中的Exo 70表达显著降低了这些细胞的侵袭和迁移能力。这些结果表明，Exo 70参与了A7 r5细胞迁移的过程。本研究对研究血管内膜增生的病理过程具有重要意义，为动脉粥样硬化、球囊血管成形术后再狭窄等心血管疾病的治疗提供了新的研究方向。
As a key subunit of the exocyst complex, Exo70 has highly conserved sequence and is widely found in yeast, mammals, and plants. In yeast, Exo70 mediates the process of exocytosis and promotes anchoring and integration of vesicles with the plasma membrane. In mammalian cells, Exo70 is involved in maintaining cell morphology, cell migration, cell connection, mRNA splicing, and other physiological processes, as well as participating in exocytosis. However, Exo70’s function in mammalian cells has yet to be fully recognized. In this paper, the expression of Exo70 and its role in cell migration were studied in a rat vascular smooth muscle cell line A7r5. Immunofluorescent analysis the expression of Exo70, α-actin, and tubulin in A7r5 cells showed a co-localization of Exo70 and α-actin, we treated the cells with cytochalasin B to depolymerize α-actin, in order to further confirm the co-localization of Exo70 and α-actin. We analyzed Exo70 co-localization with actin at the edge of migrating cells by wound-healing assay to establish whether Exo70 might play a role in cell migration. Next, we analyzed the migration and invasion ability of A7r5 cells before and after RNAi silencing through the wound healing assay and transwell assay. The mechanism of interaction between Exo70 and cytoskeleton can be clarified by the immunoprecipitation techniques and wound-healing assay. The results showed that Exo70 and α-actin were co-localized at the leading edge of migrating cells. The ability of A7r5 to undergo cell migration was decreased when Exo70 expression was silenced by RNAi. Reducing Exo70 expression in RNAi treated A7r5 cells significantly lowered the invasion and migration ability of these cells compared to the normal cells. These results indicate that Exo70 participates in the process of A7r5 cell migration. This research is importance for the study on the pathological process of vascular intimal hyperplasia, since it provides a new research direction for the treatment of cardiovascular diseases such as atherosclerosis and restenosis after balloon angioplasty.