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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 detect the frequency of loss of heterozygosity at 9 microsatellite loci associated with 4q and 8p in patients with hepatocellular carcinoma and to explore the relationship between loss of heterozygosity and clinicopathological features. Methods: Forty-five specimens of HCC were collected and tissue DNA was extracted and tested for concentration. PCR amplification was performed. Finally, the product was separated by denaturing polyacrylamide gel electrophoresis and observed for reduction or loss of the target band. Results: The total microsatellite deletion rate of nine microsatellite loci in 45 cases of informative hepatocellular carcinoma was 66.7%. Among them, the LOH rate of D8S552 was the highest (41.9%). Statistical analysis was performed by Chi-square test or Fisher’s exact test (P <0.05) Intrahepatic metastasis (P = 0.023) were statistically different. D4S415 was 22.5%, D4S3331 was 22.3%, D8S1810 was 18.8%, D4S2954 was 15.4%, D4S3030 was 15.4%, D8S1725 was 12.5%, D8S1827 was 9.8% and D8S1754 was 6.3%. CONCLUSION: Loss of heterozygosity at D4S415, D4S3331 and D8S552 loci on chromosomes 4q and 8p is an important event in the development of hepatocellular carcinoma, suggesting that there may exist potential tumor suppressor genes adjacent to it. Provide a theoretical basis.