肿瘤抑制基因HTPAP单体型构建及其与肝癌转移潜能的关系  

Haplotype construction of tumor suppressor HTPAP and its relationship with metastatic potential of hepatocellular carcinoma

作　　者：武金才[1] 陈虒 董琼珠[2] 贾户亮[2] 任宁[2] 

机构地区：[1]海南省人民医院肝胆胰外科器官移植外科,海南海口570311 [2]复旦大学附属中山医院肝肿瘤外科肝癌研究所,上海200032

出　　处：《肝胆胰外科杂志》2013年第2期126-130,135,共6页Journal of Hepatopancreatobiliary Surgery

基　　金：国家自然科学基金项目(30600589);上海市自然科学基金项目(06ZR14021);上海市青年科技启明星计划(07QA14010)

摘　　要：目的探讨肝细胞癌(HCC)患者肿瘤组织DNA的HTPAP基因单体型与肝癌转移潜能的关系。方法提取864例HCC手术样本DNA,应用焦磷酸测序行SNPs检测,并予单体型构建,分析其与肝癌转移潜能的关系。构建HTPAP启动子单体型pGL3质粒,并分别转染MHCC-97H、HepG2肝癌细胞和正常的肝Hela细胞,分析不同HTPAP启动子单体型对基因转录活性的影响。结果共发现6个SNPs位点[-1053A/G、+64G/C、+357C/G、+1648-/TAAG(无插入和插入TAAG等位基因分别以T/G示,后同)、+1838A/G、+3528C/T]。单体型构建发现HTPAP中共存在11个单体型,其中AGCTAC、GCGGGT、AGCTGC、GCGGAT四种单体型占94.75%。临床病理特征分析提示单体型GCGGGT与高转移潜能正相关(P=0.0003)。启动子序列中两个SNPs(-1053AG和+64GC)存在完全LD,故只有三种启动子单体型(-1053AA/+64GG、-1053AG/+64GC GC和-1053GG/+64CC);双荧光报告法发现三种单体型的HTPAP启动子都具有显著转录活性,其中-1053AA/+64GG单体型启动子转录活性较-1053GG/+64CC和-1053AG/+64GC单体型启动子强,差异具有统计意义(P=0.005,0.009)。结论肿瘤转移抑制基因HTPAP的GCGGGT单体型与HCC高转移潜能正相关。HTPAP启动子单体型对基因的转录活性有显著影响并与术后复发密切相关,提示HCC中HTPAP的不同SNP单体型可能是潜在侵袭转移和复发预后的预测指标。Objective To investigate the association of haplotypes of HTPAP gene with metastatic potential of hepatocellular carcinoma. Methods Single nucleotide polymorphisms and haplotypes of HTPAP gene were identi- fied in 864 patients. The association of different haplotypes with gene transcriptional activity and metastatic potential of hepatocellular carcinoma were analyzed. Results Six single nucleotide polymorphisms （-1053A/G, ＋64G/C, ＋357C/G,＋1648-/TAAG, ＋1838A/G, and＋3528C/T） were detected. Eleven haplotypes of HTPAP were identified and the 4 most frequently occurred haplotypes （AGCTAC, GCGGGT, AGCTGC, and GCGGAT） were found in 94.75% of HCCs tested. The chinical pathological characterestic analysis performed a positive correlation between GCGGGT haplotype and metastatic potential of hepatocellular carcinoma（P=0.0003）. All the three haplotypes identified in promoter sequence （-1053AA/＋64GG, -1053AG/＋64GC, and -1053GG/＋64CC） were found with high transcriptional activity, and the transcriptional activity of haplotype -1053AA/＋64GG was higher than the other two haplotypes （P=0.005, 0.009 respectively）. Conclusion GCGGGT haplotype of HTPAP gene is closely associated with metastatic potential of HCC patients. HTPAP promoter haplotypes have a significant effect on the transcriptional activity of the gene, and are closely associated with HCC recurrence. HTPAP haplotypes might be a potential predictor of metastasis and prognosis of HCC.

关 键 词：HTPAP 癌 肝细胞 单核苷酸多态性 单体型 转移 

分 类 号：R735.7[医药卫生—肿瘤]