摘要
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.
AIM: To study the specificity and significance of the expression of imprinted gene PEG10 in hepatocellular carcinoma (HCC) and to evaluate the feasibility for PEG10 as a novel molecular target of gene therapy for HCC. METHODS: The total RNA was extracted from different tumor cell lines (liver cancer HepG2, gastric cancer SGC7901, colorectal cancer Lovo, pancreatic cancer PC3, melanoma A375 and T lymphoma Jurkat cells), normal human fetal liver cell line L02, human HCC (n = 32) and the corresponding cancer-adjacent tissues (n = 32), benign liver tissues (n = 10) and peripheral blood cells (n = 10). Then the expression of PEG 10 was detected by reverse transcription polymerase chain reaction (RT-PCR). Simultaneously, AFP expression was detected in human HCC and the corresponding cancer-adjacent tissues. RESULTS: After amplification, the length of PEG10 and AFP fragment was 455 bp and 140 bp respectively. PEG10 was markedly expressed in HepG2 cells, and weakly expressed in SGC7901, PC3, Lovo cells. PEG10 expression was found negative in L02 and other tumor cell lines. The positive rates of PEG 10 expression in HCC and the corresponding tissues were 78.1 % and 0%, but the ones for AFP were 93.8% and 59.4% respectively. There was no significant difference between PEG 10 and AFP expression in HCC tissues (P>0.05), whereas the expression of AFP (19/32) was significantly higher than that of PEG10 in cancer-adjacent tissues (0/32) (X20.01,1 = 17.05, P<0.01). PEG10 wasn't detected in benign liver tissues and normal peripheral blood cells. CONCLUSION: PEG10 is more specifically expressed in HCC than AFP, which provides evidence for PEG10 as a novel molecular target of gene therapy for HCC.
出处
《世界华人消化杂志》
CAS
北大核心
2005年第12期1408-1411,共4页
World Chinese Journal of Digestology
基金
国家自然科学基金资助项目
No.30471983~~
