含三磷腺苷的Hanks平衡盐溶液处理张氏肝癌细胞诱导新去整合素金属蛋白酶相关蛋白的表达  

Expression of novel adisintegrin and metalloproteinases related protein from human Chang-liver cells treated by Hanks balaced salt solution containing adenosine triphosphate

作　　者：孙伟伟[1] 韩金红[2] 王聪睿[2] 任太芳[3] 刘瑞[1] 李端[2] 康静[2] 许重洁[2] 丰慧根[2] 

机构地区：[1]新乡医学院三全学院生命科学技术系,河南新乡453003 [2]新乡医学院生命科学技术系,河南新乡453003 [3]新乡医学院第三附属医院妇产科,河南新乡453003

出　　处：《新乡医学院学报》2012年第6期409-412,416,共5页Journal of Xinxiang Medical University

基　　金：河南省教育厅自然科学研究项目(编号:2004922070)

摘　　要：目的研究不同条件下张氏肝癌细胞中去整合素金属蛋白酶(ADAMs)及ADAM相关蛋白的表达和功能改变。方法用热激、含有三磷腺苷(ATP)的Hanks平衡盐溶液(HBSS)处理张氏肝癌细胞,Western blotting检测AD-AM相关蛋白,对处理过的细胞构建cDNA文库,用ADAMs通用抗体对文库进行免疫筛选,对阳性克隆的插入序列测序并进行体外表达,分析蛋白的性质。结果包含ATP的HBSS处理张氏肝癌细胞诱导出2个新的相对分子质量为30 000和50 000的ADAM相关蛋白,热激处理未出现相同的反应。对处理细胞构建了cDNA文库,筛选出1个新的有碱性蛋白酶活性的ADAM相关蛋白。结论含有ATP的HBSS处理张氏肝癌细胞可以诱导出1种新的ADAM相关蛋白。Objective To investigate the expression and functional alteration of adisintegrin and metalloproteinases（ADAMs） and ADAM related protein in Chang-liver cells in different cultivation surroundings. Methods Chang-liver cells were treated with heat stress and Hanks balanced salt solution（HBSS）containing adenosine triphosphate（ATP）,and ADAM related protein was dectcted by Western blotting.cDNA expression library was constructed for Chang-liver cells,then the cDNA library was screened with ADAMs common antibody.The insert sequence from positive clone was sequenced and expressed in vitro.Finally,protease activity was assayed. Results Two new relative molecular mass of 30 000 and 50 000 ADAM related proteins were induced from Chang-liver cells induced by HBSS containing ATP,but heat stress didn′t alter ADAMs expression.cDNA expression library was constructed for treated Chang-liver cells,and a new ADAM related protein possessing alkaline protease activity was confirmed in these clones. Conclusion HBSS containing ATP can induce Chang-liver cells to express one new ADAM related protein.

关 键 词：张氏肝癌细胞 去整合素金属蛋白酶 去整合素金属蛋白酶相关蛋白 

分 类 号：R735.7[医药卫生—肿瘤]