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 heal 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.%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ADAM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30000和50000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.
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