肝细胞癌变过程中磷脂酰肌醇蛋白多糖-3 mRNA扩增及临床价值

Amplification and clinical value of hepatic glypican-3 mRNA during hepatic cancerization

目的探讨磷脂酰肌醇蛋白多糖-3(GPC-3)在肝细胞性肝癌(HCC)形成过程中的表达特点及对HCC的早期诊断价值。方法以2-乙酰氨基芴(2-FAA)喂饲雄性SD大鼠诱发肝细胞癌变,观察肝组织病理学改变,并从肝组织中提取总RNA,将GPC-3mRNA以随机引物经逆转录合成GPC-3cDNA,再以巢式聚合酶链反应扩增GPC-3基因片段。结果肝细胞在癌变过程中,形态学上呈肝细胞变性、癌前病变到肝癌形成三个发展阶段,肝组织总RNA表达水平呈动态梯度表达,表现为癌变组>癌前组>变性组;肝组织GPC-3mRNA扩增阳性率在癌前组和癌变组均为100%,肝细胞变性组为83.3%,正常对照组未见阳性;肝组织GPC-3mRNA阳性率与肝总RNA表达水平,两者呈明显的正相关(r=0.475,P<0.01)。结论GPC-3参与肝细胞的癌变过程,GPC-3mRNA异常表达是HCC早期诊断有价值的分子标志物。

Objective To investigate the expression of hepatic glypican-3 （GPC-3） and its diagnotic value at early stage of hepatocellular carcinoma （HCC） formation. Methods Hepatoma models were induced with 2-fluorenyl-aeetamide （2-FAA） in male 813 rats. Morphological changes of rat livers were observed by pathological examinations （HE staining）. Total RNA was extracted. Hepatic GPC-3 gene fragments were amplified by nested RT-PCR. Results During 2-FAA inducing HCC,histological examination of hepatic cells revealed three morphological stages of granule-like degeneration （T1）, atypical hyperplasia （precancerous stage, T2） and cancer formation （T3） with a progressingly increase of hepatic total RNA,whieh was in an order of T1〈T2〈T3. The incidences of amplified fragments of GPC-3 gene were 100% in HCC, 100% in precancerous tissues, 83.8% in degeneration liver tissues, and zero in the controls, respectively. There was a closely positive correlation between GPC-3 mRNA and total RNA expression （r=0. 475, P = 0. 001 ）. Conclusion GPC-3 may participate in heptocancero-genesis, and the abnormality of GPC-3 mRNA expression could be an useful molecular marker for early diagnosis of HCC.