Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong 226001, China.
Hepatobiliary Pancreat Dis Int. 2011 Jun;10(3):289-94. doi: 10.1016/s1499-3872(11)60048-9.
Hepatocellular carcinoma (HCC) is characterized by a multi-cause, multi-stage and multi-focus process of tumor progression. Its prognosis is poor and early diagnosis is of utmost importance. This study was undertaken to investigate the dynamic expression of oncofetal antigen glypican-3 (GPC-3) and GPC-3 mRNA in hepatocarcinogenesis and to explore their early diagnostic value for HCC.
A hepatoma model was induced in male Sprague-Dawley rats with 0.05% 2-fluorenylacetamide and confirmed by hematoxylin and eosin staining and gamma-glutamyltransferase (GGT) expression. Total RNA was purified and transcribed into cDNA by reverse transcription. Fragments of the GPC-3 gene were amplified by nested RT-PCR, and confirmed by sequencing. GPC-3 was analyzed by immunohistochemistry, Western blotting or ELISA.
Positive GPC-3 expression showed as brown granule-like staining localized in the cytoplasm. Histological examination of hepatocytes revealed three morphological stages of granule-like degeneration, atypical hyperplasia (precancerous), and cancer formation, with a progressive increase of liver total RNA and GGT expression. The incidence of liver GPC-3 mRNA and GPC-3, and serum GPC-3 was 100%, 100% and 77.8% in the HCC group, 100%, 100%, and 66.7% in the precancerous group, 83.3%, 83.3%, and 38.9% in the degeneration group, and no expression in the liver or blood of the control group, respectively. There was a positive correlation between liver GPC-3 mRNA and total RNA level (r=0.475, P<0.05) or liver GPC-3 (r=1.0, P<0.001) or serum GPC-3 (r=0.994, P<0.001).
Abnormal oncofetal antigen GPC-3 and GPC-3 mRNA expression in hepatocarcinogenesis may be promising molecular markers for early diagnosis of HCC.
肝细胞癌（HCC）的特点是肿瘤进展的多病因、多阶段和多焦点过程。其预后较差，早期诊断至关重要。本研究旨在探讨胎球蛋白相关细胞外基质成分 3（GPC-3）及其 mRNA 在肝癌发生发展中的动态表达，并探讨其对 HCC 的早期诊断价值。
雄性 Sprague-Dawley 大鼠经 0.05%2-氟乙酰氨诱导建立肝癌模型，通过苏木精和伊红染色和γ-谷氨酰转移酶（GGT）表达进行确认。通过逆转录将总 RNA 纯化并转录为 cDNA。通过嵌套 RT-PCR 扩增 GPC-3 基因片段，并通过测序进行确认。通过免疫组织化学、Western blot 或 ELISA 分析 GPC-3。
阳性 GPC-3 表达表现为定位于细胞质中的棕色颗粒状染色。肝细胞组织学检查显示颗粒状变性、非典型增生（癌前病变）和癌症形成三个形态阶段，肝总 RNA 和 GGT 表达逐渐增加。HCC 组肝 GPC-3mRNA 和 GPC-3 及血清 GPC-3 的发生率分别为 100%、100%和 77.8%，癌前病变组分别为 100%、100%和 66.7%，变性组分别为 83.3%、83.3%和 38.9%，对照组肝和血中均无表达。肝 GPC-3mRNA 与总 RNA 水平（r=0.475，P<0.05）或肝 GPC-3（r=1.0，P<0.001）或血清 GPC-3（r=0.994，P<0.001）呈正相关。
肝癌发生过程中异常的胎生抗原 GPC-3 和 GPC-3mRNA 表达可能是 HCC 早期诊断有希望的分子标志物。
