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目的 :鉴定由肝细胞癌 (HCC)相关新基因LAPTM 4B编码的蛋白质并研究其生物学特性。方法 :以Westernblot、免疫组化及双向电泳鉴定新基因的表达产物 ;以免疫共沉淀等方法研究分子间的相互作用。结果 :LAPTM4B基因编码相对分子质量分别为 35× 10 3 和 2 4× 10 3 、等电点分别为 9.0 7和 4 .6 5的两种异型分子。它们在肝癌组织及肝癌细胞系的表达显著上调 ,而相对分子质量为 35× 10 3 者尤为明显 ,并与细胞分化程度相关。LAPTM4B蛋白可与整合素α 6亚单位及表皮生长因子 (EGF)受体形成信号复合物。结论 :LAPTM4B 35蛋白在人肝癌组织及细胞系过表达 ,并可能通过与细胞表面的整合素受体及EGF受体形成复合物而参与细胞外基质成分所启动的信号转导。
OBJECTIVE: To identify proteins encoded by LAPTM 4B, a novel gene associated with hepatocellular carcinoma (HCC), and to study their biological characteristics. Methods: The expression products of new genes were identified by Western blot, immunohistochemistry and two-dimensional electrophoresis. The co-immunoprecipitation method was used to study the intermolecular interactions. RESULTS: LAPTM4B gene encoded two different types of molecules with relative molecular mass of 35 × 10 3 and 24 × 10 3, respectively, with isoelectric points of 9.0 7 and 4.6. Their expression in hepatocellular carcinoma and hepatocellular carcinoma cell lines was significantly upregulated, while the relative molecular mass of 35 × 10 3 was particularly evident, and with the degree of cell differentiation related. The LAPTM4B protein forms a signaling complex with the integrin α 6 subunit and the epidermal growth factor (EGF) receptor. CONCLUSION: LAPTM4B 35 protein is overexpressed in human hepatocellular carcinoma tissues and cell lines and may participate in the signal transduction initiated by extracellular matrix components through the formation of complexes with integrin receptors and EGF receptors on the cell surface.