论文部分内容阅读
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 function of ADAMs and ADAM-related proteins in Zhang’s liver cancer cells under different conditions. Methods The liver cancer cells were treated with heat shock and Hanks balanced salt solution (ATPase) containing Hanks balanced salt solution (HBSS). Western blotting was used to detect AD-AM related proteins. The treated cells were constructed cDNA library using ADAMs universal antibody pair. The library was subjected to immunoscreening, the inserts of positive clones were sequenced and expressed in vitro, and the properties of the proteins were analyzed. Results The HBSS containing ATP treated Zhang’s hepatoma cells induced two new ADAM-related proteins with molecular weights of 30 000 and 50 000. Heat shock treatment did not show the same response. A cDNA library was constructed on the treated cells to screen out a new ADAM-related protein with alkaline protease activity. Conclusions HBsAg-containing HBSS can induce a new type of ADAM-related protein.