肝细胞癌染色体4q和8p杂合性丢失的研究

Loss of heterozygosity on Chromosome 4q and 8p in hepatocellular carcinoma

目的:检测肝细胞性肝癌患者的4q和8p两个区域上相关的9个微卫星位点的杂合性丢失频率,探讨杂合性丢失与临床病理学特征的关系。方法:选取45例信息性肝癌标本,提取组织DNA,并检测浓度,然后进行PCR扩增,最后将产物通过变性聚丙烯酰胺凝胶电泳分离并观察目的条带有无密度减少或丢失。结果:45例信息性肝癌病例的9个微卫星位点总染合性缺失率为66.7%。其中D8S552的LOH率最高为41.9%,通过χ2检验或Fisher确切概率法进行统计学分析(P<0.05),得到该位点在性别(P=0.023),血清AFP(P=0.004),有无肝内转移(P=0.023)中均有统计学差异。D4S415为22.5%,D4S3331为22.3%,D8S1810为18.8%,D4S2954为15.4%,D4S3030为15.4%;D8S1725为12.5%,D8S1827为9.8%,D8S1754为6.3%。结论:染色体4q和8p上D4S415、D4S3331、D8S552位点杂合性丢失是肝细胞性肝癌发生发展的重要事件,提示其临近部位可能存在潜在的抑癌基因;为其他肝癌相关抑癌基因的研究提供理论基础。

Objective：To study the frequency of the loss of heterozygosity of nine microsatellites located on the interrelated chromosome regions,4q and 8p in hepatocellular carcinoma（HCC）,to discuss the relationship between LOH and characteristic of clinicopathology in HCC.Methods： Tumor tissue DNA was extracted from 45 information cases which embed by paraffin and then the concentration was detected by PCR amplification.Finally the product was separated by D-PAGE in order to observe whether there＇s a decrease of the density or deletion of the target bands in informative cases.Results： All 45 cases informative liver cancer patients were tested and the total rate of LOH at the 9 microsatellite loci was 66.7%.Among these microsatellite loci,D8S552 had a highest rate of LOH,which was 41.9%,and there were significant differences in both gender（P=0.023）,serum AFP（P=0.004）,intrahepatic metastasis conditions（P=0.023）.The rate of LOH at D4S415 was 22.5%,while D4S3331 was 22.3%,D8S1810 was 18.8%,D4S2954 was 15.4%,D4S3030 was 15.4%;D8S1725 was 12.5%,D8S1827 was 9.8%,D8S1754 was 6.3%.Conclusion： The loss of heterozygosity at D4S415,D4S3331,D8S552 locus are important events in the development of hepatocellular carcinoma.It suggests that there may be potential tumor suppressor genes nearby the position,as well as other tumor suppressor genes in hepatocellular carcinoma.And it also provides a theoretical basis for study of other tumor suppressor genes related to liver cancer.