A Serological Marker for Hepatocellular Carcinoma

描述(由申请人提供)：编码磷脂酶A2组13(PLA2G13)的基因在肝细胞癌(HCC)组织中的表达显著上调，相对于邻近的非恶性肝脏。在对公共基因表达数据库的搜索中，只发现了一个包含PLA2G13编码序列的cDNA文库，这表明该基因很少表达或表达水平很低。由于PLA2G13是一种分泌型蛋白，因此有可能成为肝癌的血清学标志物。这项建议的重点是通过开发基于抗体的PLA2G13检测来检验这一假设，以便确定患者血清中的肝癌水平。在该项目的第一阶段，将生产针对三种不同合成PLA2G13多肽的兔多克隆抗体。针对重组人PLA2G13(RhPLA2G13)产生的多克隆抗体也将从学术合作者那里获得。以重组人PLA2G13为标准，用Western Blot和夹心ELISA法检测肝细胞癌患者和正常人血清中该蛋白的存在。PLA2G13的相对水平将与其他参数相关，包括甲胎蛋白水平和肿瘤病理。作为这一项目阶段的先决条件，将获得机构审查委员会的批准，以获取更多的血清样本，以更好地表征PLA2G13作为肝细胞癌的血清学标志。

DESCRIPTION (provided by applicant): Expression of the gene encoding phospholipase A2 group 13 (PLA2G13) was found to be significantly upregulated in hepatocellular carcinoma (HCC) tissue relative to adjacent nonmalignant liver. In a search of public gene expression databases, only one cDNA library was found to contain the sequence encoding PLA2G13 suggesting that the gene is rarely expressed or expressed at very low levels. Since PLA2G13 is a secreted protein, it has potential as a serological marker of HCC. The focus of this proposal is to test this hypothesis by developing antibody based assays for PLA2G13 in order to determine HCC levels in patient sera. In phase I of this project, rabbit polyclonal antibodies to three different synthetic PLA2G13 peptides will be produced. Polyclonal antibodies generated against recombinant human PLA2G13 (rhPLA2G13) will also be obtained from academic collaborators. Using rhPLA2G13 as a standard, serum from HCC patients and normal individuals will be assayed by Western Blot and sandwich ELISA for the presence of the protein. Relative levels of PLA2G13 will be correlated with other parameters including alpha-fetoprotein levels and tumor pathology. As a prerequisite for phase of this project, Institutional Review Board approval will be obtained to acquire more serum samples to better characterize PLA2G13 as a serological marker for HCC.