Effects of CASK down-regulation on the proliferation of ECV-304 cells

陈建; 苏踊跃; 梁光萍; 陈渝; 罗向东; 杨宗城

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Effects of CASK down-regulation on the proliferation of ECV-304 cells

更新时间：2026-01-09

- Effects of CASK down-regulation on the proliferation of ECV-304 cells
- Journal of Army Medical University Vol. 31, Issue 12, Pages: 1144-1146(2009)
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  CLC： R322.12;R329.28;R345
- Published：2009
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陈建,苏踊跃,梁光萍,陈渝,罗向东,杨宗城.CASK下调表达对人血管内皮细胞ECV-304增殖能力的影响[J].陆军军医大学学报(原第三军医大学学报),2009,31(12):1144-1146.

　CHEN Jian,SU Yong-yue,LIANG Guang-ping,CHEN Yu,LUO Xiang-dong,YANG Zong-cheng.Effects of CASK down-regulation on the proliferation of ECV-304 cells[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(12):1144-1146.
陈建,苏踊跃,梁光萍,陈渝,罗向东,杨宗城.CASK下调表达对人血管内皮细胞ECV-304增殖能力的影响[J].陆军军医大学学报(原第三军医大学学报),2009,31(12):1144-1146. DOI：

　CHEN Jian,SU Yong-yue,LIANG Guang-ping,CHEN Yu,LUO Xiang-dong,YANG Zong-cheng.Effects of CASK down-regulation on the proliferation of ECV-304 cells[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(12):1144-1146. DOI：

摘要

目的建立CASK下调表达细胞株，研究钙/钙调蛋白依赖性丝氨酸蛋白激酶（calcium

calmodulin-associated serine/threonine kinase

CASK）下调表达对ECV-304细胞增殖能力的影响。方法将包含针对CASK基因干扰siRNA片段的pGenesil-1-hCASK重组质粒转染人ECV-304细胞株，经G418加压筛选，成功筛选出CASK下调表达的细胞株。用蛋白免疫印迹法鉴定转染细胞中CASK基因的表达水平。采用流式细胞技术、MTT法观察CASK下调表达对ECV-304细胞增殖能力的变化。结果成功筛选出CASK下调表达ECV-304细胞株（siCASK细胞株），这些细胞在传代过程中绿色荧光蛋白能维持表达，细胞株中转染阳性率在90%以上；经蛋白免疫印迹法检测CASK表达明显降低； siCASK细胞株中G

细胞比率下降，G

M期细胞比率明显上升，增殖指数明显提高，生长曲线左移。结论成功建立了CASK下调表达的ECV-304细胞株，CASK下调表达可导致细胞增殖能力增强。

Abstract

Objective To establish calcium

calmodulin-associated serine/threonine kinase (CASK) down-regulation ECV-304 cell line and to study the effects of CASK down-regulation on the proliferation of ECV-304 cells. Methods The recombinant pGenesil-1-hCASK plasmids containing siRNA targeting CASK gene were transfected into ECV-304 cells. The stable cell strain was screened by adding G418 into cell culture medium. The protein expression levels of CASK gene were detected by Western Blotting. Flow cytometry and MTT method were used to investigate the effects of CASK down-regul

ation on the proliferation of the transfected cells. Results CASK down-regulation cell strain

siCASK cell line

was successfully screened. These cells could maintain over 90% green fluorescent protein expression. The decreased expression of CASK could lower but enhance the cell population at G

phase but G

M phase and lead to left-shift of the cell growth curve. Conclusion CASK down-regulation cell strain was successfully constructed and CASK down-regulation could enhance the cell proliferation.

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