Eukaryotic expression cloning with an antimetastatic monoclonal antibody identifies a tetraspanin (PETA-3/CD151) as an effector of human tumor cell migration and metastasis.
Eukaryotic expression cloning with an antimetastatic monoclonal antibody identifies a tetraspanin (PETA-3/CD151) as an effector of human tumor cell migration and metastasis.
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发表时间：
1999-08
期刊：
影响因子：
11.2
通讯作者：
J. Testa;P. Brooks;J. Lin;J. Quigley
中科院分区：
文献类型：
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作者：
J. Testa;P. Brooks;J. Lin;J. Quigley
发现通过消减免疫产生的单克隆抗体（mAb）50-6特异性抑制人表皮样癌细胞系HEp-3的体内转移。通过改良的真核表达克隆方案从HEp-3文库中分离mAb 50-6的同源抗原的cDNA。序列分析鉴定抗原为PETA-3/CD 151，最近描述的四跨膜蛋白家族的成员。克隆的抗原也被先前描述的抗转移抗体mAb 1A 5识别。mAb对HEp-3转移的抑制不能归因于抗体对体外或体内肿瘤细胞生长的任何影响。相反，抗体似乎抑制了转移灶形成的早期步骤。在趋化性测定中，HEp-3迁移被两种抗体阻断。HeLa细胞转染和过表达PETA-3/CD 151比对照转染表达很少的CD 151迁移。HeLa迁移的增加可通过mAb 50-6和mAb 1A 5两者来实现。PETA-3似乎不参与细胞附着，因为粘附与PETA-3表达水平无关，并且不受mAb 50-6或mAb 1A 5的影响。PETA-3介导细胞迁移的能力表明了这种蛋白质可能影响转移的机制。这些数据确定PETA-3/CD 151是四跨膜蛋白家族的第一个成员，与转移的阳性效应物相关。
A monoclonal antibody (mAb), 50-6, generated by subtractive immunization, was found to specifically inhibit in vivo metastasis of a human epidermoid carcinoma cell line, HEp-3. The cDNA of the cognate antigen of mAb 50-6 was isolated by a modified eukaryotic expression cloning protocol from a HEp-3 library. Sequence analysis identified the antigen as PETA-3/CD151, a recently described member of the tetraspanin family of proteins. The cloned antigen was also recognized by a previously described antimetastatic antibody, mAb 1A5. Inhibition of HEp-3 metastasis by the mAbs could not be attributed to any effect of the antibodies on tumor cell growth in vitro or in vivo. Rather, the antibodies appeared to inhibit an early step in the formation of metastatic foci. In a chemotaxis assay, HEp-3 migration was blocked by both antibodies. HeLa cells transfected with and overexpressing PETA-3/CD151 were more migratory than control transfectants expressing little CD151. The increase in HeLa migration was inhibitable by both mAb 50-6 and mAb 1A5. PETA-3 appears not to be involved in cell attachment because adhesion did not correlate with levels of PETA-3 expression and was unaffected by mAb 50-6 or mAb 1A5. The ability of PETA-3 to mediate cell migration suggests a mechanism by which this protein may influence metastasis. These data identify PETA-3/CD151 as the first member of the tetraspanin family to be linked as a positive effector of metastasis.