Expression of the extracellular domain of OB-cadherin as an Fc fusion protein using bicistronic retroviral expression vector
Expression of the extracellular domain of OB-cadherin as an Fc fusion protein using bicistronic retroviral expression vector
复制标题
DOI：
10.1016/j.pep.2008.06.007
复制
发表时间：
2008-10-01
影响因子：
1.6
通讯作者：
Lin, Sue-Hwa
中科院分区：
文献类型：
--
作者：
Lira, Cristina B. B.;Chu, Khoi;Lin, Sue-Hwa
成骨细胞钙粘蛋白（OB-cadherin，也称为钙粘蛋白-11）是一种主要在成骨细胞中表达的钙依赖性嗜同性细胞粘附分子。OB-钙粘蛋白在前列腺癌中表达，并可能参与转移性前列腺癌细胞向骨的归巢。OB-cadherin的胞外结构域可用于抑制前列腺癌细胞与成骨细胞之间的粘附。在这份报告中，我们描述的表达OB-钙粘蛋白的胞外结构域的Fc融合蛋白（OB-CAD-Fc）在人胚肾293 FT细胞使用双顺反子逆转录病毒载体。通过双顺反子载体共表达GFP和OB-CAD-Fc允许通过荧光激活细胞分选富集表达OB-CAD-Fc的细胞。将重组OB-CAD-Fc蛋白分泌到细胞培养基中，并使用固定化蛋白A-亲和层析从11的条件培养基中纯化约0.85mg纯化的OB-CAD-Fc蛋白。纯化的CB-CAD-Fc具有生物活性，因为它支持用OB-钙粘蛋白转导的PC 3细胞和L细胞的粘附。OBCAD-Fc的可用性提供了测试OB-CAD-Fc是否可用于抑制OB-钙粘蛋白介导的前列腺癌体内骨转移或产生抑制前列腺癌细胞与成骨细胞之间粘附的抗体的机会。(c)2008年爱思唯尔公司All rights reserved.
Osteoblast cadherin (OB-cadherin, also known as cadherin-11) is a Ca2+-dependent homophilic cell adhesion molecule that is expressed mainly in osteoblasts. OB-cadherin is expressed in prostate cancer and may be involved in the homing of metastatic prostate cancer cells to bone. The extracellular domain of OB-cadherin may be used to inhibit the adhesion between prostate cancer cells and osteoblasts. In this report, we describe the expression of the extracellular domain of OB-cadherin as an Fc fusion protein (OB-CAD-Fc) in human embryonic kidney 293FT cells using a bicistronic retroviral vector. Coexpression of GFP and OB-CAD-Fc through the bicistronic vector permitted enrichment of OB-CAD-Fc-expressing cells by fluorescence-activated cell sorting. Recombinant OB-CAD-Fc proteins were secreted into cell medium, and about 0.85 mg of purified OB-CAD-Fc protein was purified from I I of the conditioned medium using immobilized protein A-affinity chromatography. The purified CB-CAD-Fc was biologically active because it supported the adhesion of PC3 cells and L cells transduced with OB-cadherin. The availability of OBCAD-Fc offers opportunities to test whether OB-CAD-Fc can be used to inhibit OB-cadherin-mediated prostate cancer bone metastasis in vivo or to generate antibodies for inhibiting the adhesion between prostate cancer cells and osteoblasts. (c) 2008 Elsevier Inc. All rights reserved.