EJ细胞CD44基因RNAi对单抗KMP1和细胞功能的影响

基础医学与临床 ›› 2012, Vol. 32 ›› Issue (3): 313-316.

EJ细胞CD44基因RNAi对单抗KMP1和细胞功能的影响

丁明霞,李翀,左毅刚,王剑松

昆明医学院第二附属医院泌尿外科

收稿日期:2011-04-21 修回日期:2011-12-23 出版日期:2012-03-05 发布日期:2012-02-27
通讯作者: 王剑松 E-mail:jiansongwang@yohoo.com
基金资助:
云南省科技厅高层次人才培引工程经费资助项目

CD44 knockdown decreases immuno- bounding to the monoclonal antibody KMP1 and inhibits the growth and metastasis of EJ cells

Received:2011-04-21 Revised:2011-12-23 Online:2012-03-05 Published:2012-02-27

摘要/Abstract

摘要： 研究EJ细胞中CD44基因小RNA干扰后细胞功能的变化和单抗KMP1功能的改变。方法小RNA干扰的方法抑制EJ细胞中CD44基因表达，Western blot和流式细胞仪检测EJ细胞与CD44特异性单抗KMP1的结合功能变化，细胞划痕实验和Boyden小室研究shCD44－EJ细胞的迁移功能改变，[3H]掺入法检测shCD44－EJ细胞的增殖能力变化。结果　KMP1与shCD44－EJ细胞结合功能下降，shCD44－EJ细胞的迁移能力降低约50%，增殖能力也下降。结论　特异性沉默CD44可降低EJ细胞特异性单抗KMP1的结合功能，并抑制EJ细胞的迁移和增殖能力。

关键词: 膀胱癌, EJ细胞, CD44, RNA干扰

Abstract: [Abstract] Objective To investigate the effect of CD44 targeted RNA interference (RNAi) on the growth and Metastasis of human bladder carcinoma EJ cells. Methods We transfected bladder cancer cell line EJ with well-designed CD44 siRNA. Flow cytometry and western blot analysis were used to assess the CD44 immuno- bounding to the monoclonal antibody KMP1. Wound healing assay and Boyden chamber were used to investigate the change of shCD44－EJ cell’s migration. Proliferation of cells was measured using a [3H]-thymidine incorporation assay. Results CD44 knockdown extremely decreased KMP1 staining signal compared with the EJ cells transfected by pSuper-puro empty vector control，Both flow cytometry and western blot analysis revealed a remarkable decrease of CD44 expression in shCD44－EJ cells，but not in pSuper-puro vector transfected cells. CD44 silencing also significantly inhibited cell migration through wound healing assay. The modified Boyden chamber assay showed similar results also. Conclusions CD44 silencing can effectively decrease EJ cells immuno- bounding to the monoclonal antibody KMP1 and inhibit the migration and the proliferation of EJ cells in vitro.

Key words: bladder carcinoma, EJ cells, CD44, RNA interference

中图分类号:

丁明霞李翀左毅刚王剑松. EJ细胞CD44基因RNAi对单抗KMP1和细胞功能的影响[J]. 基础医学与临床, 2012, 32(3): 313-316.

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