A Serological Marker for Hepatocellular Carcinoma

描述（由申请人提供）：发现编码磷脂酶A2组13（PLA2G13）的基因的表达在肝细胞癌（HCC）组织中相对于邻近的非恶性肝脏显著上调。在公共基因表达数据库的搜索中，仅发现一个cDNA文库含有编码PLA2G13的序列，表明该基因很少表达或以非常低的水平表达。由于PLA2G13是一种分泌性蛋白，它有可能作为HCC的血清学标志物。该提案的重点是通过开发基于抗体的PLA2G13测定来测试该假设，以确定患者血清中的HCC水平。在该项目的第一阶段，将产生针对三种不同合成PLA2G13肽的兔多克隆抗体。还将从学术合作者处获得针对重组人PLA2G13（rhPLA2G13）生成的多克隆抗体。使用rhPLA2G13作为标准品，通过Western印迹和夹心ELISA测定来自HCC患者和正常个体的血清中蛋白质的存在。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学。作为本项目阶段的先决条件，将获得机构审查委员会的批准，以获得更多的血清样本，以更好地表征PLA2G13作为HCC的血清学标志物。

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.