敲除Wdr1抑制小鼠原代血管平滑肌细胞的迁移和增殖

doi:10.13560/j.cnki.biotech.bull.1985.2018-0578

摘要/Abstract

摘要： 细胞骨架肌动蛋白在细胞增殖、迁移等生物学过程中发挥重要作用,然而 WDR1 作为肌动蛋白解聚的主要辅助因子,其在平滑肌细胞中的作用尚无报道。构建 Wdr1 条件性诱导敲除小鼠模型,进而分离小鼠原代主动脉平滑肌细胞,诱导敲除 Wdr1,检测平滑肌细胞的形态、增殖和迁移情况。PCR 结果显示,Wdr1^f/f;ERT2Cre 小鼠模型构建成功。免疫荧光结果表明：在前人基础上改进的分离方法能够高效分离原代平滑肌细胞。细胞形态观察结果显示：Wdr1 敲除后对细胞的形态有显著性影响。同时,划痕实验以及 CCK8 检测结果也表明：Wdr1 的条件性诱导性敲除可明显抑制平滑肌细胞的迁移和增殖。Wdr1 敲除后平滑肌细胞的形态以及细胞生物学过程的改变提示其与血管疾病的发生和发展有紧密联系,这对揭示血管病理生理过程有重要意义。

关键词: 条件性诱导敲除小鼠, WDR1, 平滑肌细胞, 增殖, 迁移

Abstract: Cytoskeletal actin plays an important role in cell proliferation,migration,and other biological processes. However,the role of WDR1,a major cofactor for actin depolymerizing,has not been reported in smooth muscle cells（SMCs）. Inducible Wdr1 conditional knockout mouse model was constructed,and then sacrificed to isolate mouse primary vascular smooth muscle cells（VSMCs）and induced to detect the morphology,proliferation,and migration. The PCR results suggested that Wdr1^f/f;ERT2Cre mouse model was successfully constructed. The results of immunofluorescence revealed that the improved separation method can effectively separate primary smooth muscle cells. A significant effect on cell morphology after Wdr1 knockout was observed. Concurrently,the scratch and CCK8 test results also showed that Wdr1 conditional knockout obviously inhibited the migration and proliferation of SMCs. The changes of smooth muscle cell morphology and cell biological processes after Wdr1 knockout indicate that it is closely related to the occurrence and development of vascular diseases. It is of great significance to uncover the pathophysiological process of blood vessels.

Key words: conditional induced knockout mouse, WDR1, smooth muscle cells, proliferation, migration

袁白银, 胡继盛, 刘钟颖, 黄霞. 敲除Wdr1抑制小鼠原代血管平滑肌细胞的迁移和增殖[J]. 生物技术通报, 2018, 34(12): 179-185.

YUAN Bai-yin, HU Ji-sheng, LIU Zhong-ying, HUANG Xia. Wdr1 Knockout Inhibits Migration and Proliferation of Mouse Primary Vascular Smooth Muscle Cells[J]. Biotechnology Bulletin, 2018, 34(12): 179-185.

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