肾细胞癌中染色体3p、9p、14q微卫星不稳定及杂合性缺失分析

Microsatellite instability and loss of heterozygosity on chromosome 3p,9p and 14q in renal cell carcinoma

目的:探讨肾细胞癌患者的微卫星不稳定(MSI)和杂合性缺失(LOH)的发生频率,并同时研究染色体微卫星变异与肾细胞癌临床病理特征之间的关系。方法:选取位于染色体3p、9p、14q上的共计12个微卫星多态性标记,采用PCR-中性聚丙烯酰胺凝胶电泳-EB染色和测序的方法对31例肾细胞癌患者的原发病灶和配对的转移病灶进行MSI和LOH分析。结果:MSI发生频率可达61.3%。LOH频率可达54.8%,其中以D9S168的MSI发生率最高为32.3%。LOH发生率最高的为D3S1289。达21.4%。经统计学检验结果显示,MSI的发生频率与肾细胞癌的TNM分期具有相关性(P<0.05).但MSI和LOH与肾细胞癌患者的性别、年龄和病理类型以及转移均无关(P>0.05)。结论:除D3S1566外的11个微卫星标记的MSI和LOH可能是一个增加肾癌易感性的危险因素,D9S168、D3S1289是肾细胞癌中敏感的检测位点,在D9S168和D3S1289附近可能存在与肾细胞癌有关的癌基因/抑癌基因,从而影响肾癌的发生和发展。

Objective： To investigate frequencies of microsatellite instability （MSI） and loss of heterozygosity （LOH） in renal cell carcinoma （RCC）,and to discuss the relationship of clinicopathological characteristics of RCC with MSI and LOH. Methods： Twelve microsatellite markers located at chromosomes 3p, 9p and 14q were selected to investigate microsatellite alterations （MSI and LOH） in 31 RCC specimens and their paired metastasis specimens by polymerase chain reactionpolyacrylamide gel electrophoresis-ethylene dibromide （PCR-PAGE-EB） staining and sequencing. Results： The frequency of MSI could reached 61.3% and that of LOH could reach 54.8%. The highest frequency of MSI was at locus of D9S168 （32.3%） ; the highest frequency of LOH was at locus of D3S1289 （21.4%）. No correlation was found between MSI or LOH and the patients＇ age,sex,pathology type and metastastis,except that MSI was correlated with TNM stage of RCC （P〈0.05）. Conclusion： MSI and LOH of 12 microsatellite markers, except for D3S1566, might be risk factors for RCC. D9S168 and D3S1289 are two sensitive loci in RCC, and they might be close to RCC-associated oncogenes or anti-oncogenes, which may influence the development and progression of RCC.