Transfection of MCF-7 carcinoma cells with human integrin alpha7 cDNA promotes adhesion to laminin.
Transfection of MCF-7 carcinoma cells with human integrin alpha7 cDNA promotes adhesion to laminin.
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用人整合素 α7 cDNA 转染 MCF-7 癌细胞可促进层粘连蛋白的粘附。
DOI：
10.1006/abbi.2000.2134
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发表时间：
2001
影响因子：
3.9
通讯作者：
R. Kramer
中科院分区：
文献类型：
--
作者：
I. Vizirianakis;C. Yao;Y. Chen;B. Ziober;A. Tsiftsoglou;R. Kramer
层粘连蛋白结合α 7 β 1整联蛋白受体在骨骼肌和心肌中高度表达，并被认为是肌源性细胞迁移和分化过程中的关键分子。整联蛋白α 7亚基的缺失导致整联蛋白α 7缺失小鼠的一种形式的肌营养不良，而α 7基因中的特定突变与人类的先天性肌病相关。为了更详细地研究整合素α 7在人类相关肌肉疾病中的潜在作用，我们通过RT-PCR从人骨骼肌mRNA中克隆α 7 cDNA，然后通过转染在包括MCF-7、COS-7和NIH 3 T3细胞在内的几种细胞系中表达全长人整合素α 7 cDNA。分离的cDNA对应于人α 7 X2 B选择性剪接形式。通过转染嵌合人/小鼠α 7 cDNA构建体进一步证实人α 7的表达。为了证明表达的人α 7的功能性，用转染的MCF-7细胞进行的粘附实验已经证实了人α 7与层粘连蛋白的特异性结合。此外，产生抗人α 7的细胞外结构域的小鼠多克隆和单克隆抗体，并用于通过流式细胞术分析用全长人α 7 cDNA转染的MCF-7和NIH 3 T3细胞。这些结果首次显示了功能性全长人α 7 cDNA的外源表达，以及针对人α 7细胞外结构域的单克隆抗体的开发。开发的抗体将用于进一步分析涉及α 7功能障碍的人类疾病，并促进肌肉干细胞（卫星细胞）的分离，从而扩大人类疾病的遗传修饰移植治疗的机会。
The laminin-binding alpha7beta1 integrin receptor is highly expressed by skeletal and cardiac muscles, and has been suggested to be a crucial molecule during myogenic cell migration and differentiation. Absence of integrin alpha7 subunit contributes to a form of muscular dystrophy in integrin alpha7 null mice, whereas specific mutations in the alpha7 gene are associated in humans with congenital myopathy. To examine in more detail the potential role of integrin alpha7 in human-related muscular disorders, we cloned alpha7 cDNA by RT-PCR from human skeletal muscle mRNA and then expressed the full-length human integrin alpha7 cDNA by transfection in several cell lines including MCF-7, COS-7, and NIH3T3 cells. The isolated cDNA corresponds to the human alpha7X2B alternative splice form. Expression of human alpha7 was further confirmed by transfection of chimeric human/mouse alpha7 cDNA constructs. To demonstrate the functionality of expressed human alpha7, adhesion experiments with transfected MCF-7 cells have confirmed the specific binding of human alpha7 to laminin. In addition, mouse polyclonal and monoclonal antibodies were generated against the extracellular domain of human alpha7 and used to analyze by flow cytometry MCF-7 and NIH3T3 cells transfected with the full-length of human alpha7 cDNA. These results show for the first time the exogenous expression of functional full-length human alpha7 cDNA, as well as the development of monoclonal antibodies against the human alpha7 extracellular domain. Antibodies developed will be useful for further analysis of human disorders involving alpha7 dysfunction and facilitate isolation of muscle stem cells (satellite cells) and thereby expand the opportunities for genetically modified transplantation treatment of human disease.
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影响因子：
4
作者：
Z. Bao;M. Lakonishok;S. Kaufman;A. Horwitz
通讯作者：
Z. Bao;M. Lakonishok;S. Kaufman;A. Horwitz
DOI：
--
发表时间：
1989-09
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
R. Kramer;K. Mcdonald;M. Vu
通讯作者：
R. Kramer;K. Mcdonald;M. Vu
影响因子：
7.5
作者：
Frisch, SM;Ruoslahti, E
通讯作者：
Ruoslahti, E
影响因子：
7.5
作者：
Hemler, ME
通讯作者：
Hemler, ME
α7 整联蛋白受体的层粘连蛋白结合活性由胞外域中发育调节的剪接定义。
影响因子：
3.3
作者：
Ziober,BL;Chen,Y;Kramer,RH
通讯作者：
Kramer,RH