贵州医科大学学报

2011, v.36;No.150(03) 235-237+241

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钙激活中性蛋白酶在肝癌细胞迁移能力中的作用
A Research on the Function of Calcium-activated Neutral Protease in Metastasis of Hepatocarcinoma Cell

唐佳艳;洪阳;向敏;郑洋;王艳;胡晓霞;潘娅;王旭东;周力;陈腾祥;
TANG Jiayan1,2,HONG Yang3,XIANG Min4,ZHENG Yang1,2,WANG Yan2,HU Xiaoxia2,PAN Ya2,WANG Xudong2,ZHOU Li1,CHEN Tengxiang2 (1.Division of Gastroenterology,the Affiliated Hospital of Guiyang Medical College,Guiyang 550004,Guizhou,China;2.Department of Physiology,Guiyang Medical College,Guiyang 550004,Guizhou,China;3.Public Health Bureau of Guiyang,Guiyang 550014,Guizhou,China;4.Division of Pediatrics,the Affiliated Hospital of Guiyang Medical College,Guiyang 550004,Guizhou,China)

摘要(Abstract):

目的:观察不同转移能力肝癌细胞中钙激活中性蛋白酶(calpain)的表达情况,利用抑制剂抑制cal-pain活性,评价calpain对肝癌细胞迁移能力的影响。方法:用蛋白质印迹(Western blotting)检测不同转移能力肝癌细胞或肝细胞中calpain-1、calpain-2和Src的表达和酶解情况;用划痕法观察calpain-2抑制剂对高转移性肝癌细胞(MHCC97-H)迁移能力的影响。结果:Western blot结果显示,不同肝细胞系中calpain-1、calpain-2及Src的表达水平和剪切成为低分子量(LMW)活性形式的程度不一样,其中calpain-2和Src的表达与肝癌细胞的迁移能力有相关性,calpain-2抑制剂明显下调MHCC97-H细胞中calpain-2和Src的表达,减少其被剪切的程度;划痕实验结果显示,calpain-2抑制剂能明显减少MHCC97-H细胞向划痕区域迁移,而且抑制剂浓度越大,细胞迁移能力被抑制的程度就越重。结论:calpain-2的表达上调和活性增强可通过激活Src增加肝癌细胞的迁移能力。
Objective:To evaluate the effect of calcium-activated neutral protease(calpain) on metastasis of hepatocarcinoma cell.Methods:Western blotting assay was used to detect the expression and zymohydrolysis of calpain-1,calpain-2 and Src in hepatoma cell or hepatocyte cell lines with different metastasis ability,while wound healing assay was used to detect the effect of calpain-2 inhibitor on the migration ability of MHCC97-H(hepatocarcinoma cell with high metastasis ability).Results:As Western blot result showing,the expression and the active low molecular weight(LMW) forms of calpain-1,calpain-2 and Src from zymohydrolysis were not consistent in different hepatic cell lines,in which the alteration model of calpain-2 and Src were related with the metastasis ability except calpain-1.In MHCC97-H cell line,the expression level and active LMW forms of calpain-2 and Src were reduced by calpain-2 inhibitor.Consistently,with the inhibitor of calpain-2,the migration of MHCC97-H was reduced,and the higher the inhibitor concentration was,the stronger the inhibition of cell migration ability was.Conclusions:In calpain family,the up-regulation of calpain-2 expression and increase of its activity might increase the migration ability of hepatoma cells by activate Src.

关键词(KeyWords): 肝癌细胞;钙激活中性蛋白酶;细胞迁移;印迹法,蛋白质
hepatocarcinoma cell;calcium-activated neutral proteases;cell migration;blotting,western

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金资助项目(NO.81060176);; 贵州省科技厅自然基金项目[黔科合J字(2008)2280];; 贵阳市科技局社会发展公关计划项目[筑科农字(2010)1号]

作者(Author): 唐佳艳;洪阳;向敏;郑洋;王艳;胡晓霞;潘娅;王旭东;周力;陈腾祥;
TANG Jiayan1,2,HONG Yang3,XIANG Min4,ZHENG Yang1,2,WANG Yan2,HU Xiaoxia2,PAN Ya2,WANG Xudong2,ZHOU Li1,CHEN Tengxiang2 (1.Division of Gastroenterology,the Affiliated Hospital of Guiyang Medical College,Guiyang 550004,Guizhou,China;2.Department of Physiology,Guiyang Medical College,Guiyang 550004,Guizhou,China;3.Public Health Bureau of Guiyang,Guiyang 550014,Guizhou,China;4.Division of Pediatrics,the Affiliated Hospital of Guiyang Medical College,Guiyang 550004,Guizhou,China)

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DOI: 10.19367/j.cnki.1000-2707.2011.03.005

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