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 identified 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.
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