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目的:研究PEG10在肝癌组织中表达的特异性,为其作为一个潜在的肝癌基因治疗的新的分子靶点提供实验依据.方法:从来自不同器官组织的肿瘤细胞系(人肝癌细胞株HepG2、人胃癌细胞株SGC7901、人结肠癌细胞株Lovo、人胰腺癌细胞株PC3、人黑色素瘤细胞株A375、人T淋巴瘤细胞株Jurkat)、正常人胎肝细胞株L02、32例肝癌患者的肝癌组织、癌旁组织、10例良性肝病患者肝组织、10例正常人外周血单个核细胞中抽提总RNA,经RT逆转录合成cDNA,再以PCR方法检测PEG10的表达.同时,肝癌组织和癌旁组织标本经RT-PCR检测AFP的表达.结果:经RT-PCR扩增的PEG10基因片段为455bp,AFP基因的扩增片段为140bp,与原设计一致;PEG10在人肝癌细胞株HepG2中明显表达,人胃癌细胞株SGK7901、人结肠癌细胞株Lovo、人胰腺癌细胞株PC3中弱表达,而正常人胚胎肝细胞株LO2和其他肿瘤细胞株(人T淋巴细胞瘤、人黑色素瘤)中表达均为阴性雇32例肝癌及相应癌旁组织中,PEG10的表达阳性率分别为78.1%和0%;而AFP基因的阳性率分别为93.8%和59.4%.经McNemar检验,显示PEG10基因在肝癌组织中表达的敏感性与AFP表达敏感性之间无显著性差异(X20.01,1=1.78,P>0.05);而癌旁组织中PEG10表达率(0/32)明显低于AFP表达率(19/32)(X20.01,1=17.05,P<0.01).另外10例良性肝病患者(肝硬化4例,自身免疫性肝病2例,肝血管瘤2例,丙型肝炎1例,血色素病1例)肝组织标本及正常人的外周血细胞进行PEG10基因检测,结果均为阴性.结论:PEG10的表达不但具有比AFP更高的肝癌组织特异性,而且具有相对器官组织特异性.本实验为PEG10作为一个新的肝癌标志物和基因治疗的分子靶点提供了实验依据.
OBJECTIVE: To study the specificity of PEG10 in hepatocellular carcinoma (HCC) and to provide experimental evidence for its potential as a new molecular target for hepatocellular carcinoma gene therapy.Methods: Tumor cell lines from different organs and tissues (HepG2, Human gastric cancer cell line SGC7901, human colon cancer cell line Lovo, human pancreatic cancer cell line PC3, human melanoma cell line A375, human T lymphoma cell line Jurkat), normal human fetal hepatocyte cell line L02, and 32 liver cancer patients Total RNA was extracted from peripheral blood mononuclear cells from 10 patients with benign liver disease, 10 normal liver tissues, and reverse transcribed into cDNA by reverse transcription polymerase chain reaction (RT-PCR), and the expression of PEG10 was detected by PCR.At the same time, The expression of AFP was detected by RT-PCR.Results: The PEG10 gene fragment amplified by RT-PCR was 455bp and the AFP gene amplified fragment was 140bp, consistent with the original design; PEG10 in human hepatocellular carcinoma cell line HepG2 Obviously, the human gastric cancer cell line SGK7901, the human colon cancer cell line Lovo and the human pancreatic cancer cell line PC3 were weakly expressed, whereas the normal human embryonic liver cell line LO2 and other tumor cell lines (human T-cell lymphoma, human melanin ) Were negative hired 32 cases of liver cancer and corresponding paracancerous tissues, the positive rate of PEG10 expression were 78.1% and 0%, respectively, while the positive rate of AFP gene was 93.8% and 59.4%, respectively.The McNemar test showed that PEG10 There was no significant difference between the sensitivity of AFP expression and the sensitivity of AFP expression (X20.01,1 = 1.78, P> 0.05), while the expression of PEG10 in adjacent tissues was significantly lower than that of AFP AFP expression rate (19/32) (X20.01,1 = 17.05, P <0.01) .Other 10 patients with benign liver disease (cirrhosis in 4 cases, 2 cases of autoimmune liver disease, 2 cases of hepatic hemangioma, hepatitis C 1 case, hemochromatosis in 1 case), liver tissue samples and normal human peripheral blood cells for PEG10 gene detection, the results were negative.Conclusion: PEG10 expression not only has a higher specificity than HCC liver cancer tissue, and with relative organ tissue specificity This experiment provides the experimental basis for PEG10 as a new molecular marker of liver cancer and gene therapy.