肾透明细胞癌多中心病灶单克隆、多克隆起源并存的研究论证  

Evidence of monoclonal and polyclonal origin coexisting for multifocal clear cell renal cell carcinoma

作　　者：王增增[1] 李泉林[2] 王一明[3] 

机构地区：[1]天津市北辰医院泌尿外科,天津300400 [2]大连医科大学附属第一医院泌尿外科 [3]辽宁医学院附属第三医院泌尿外科

出　　处：《临床泌尿外科杂志》2015年第8期685-689,共5页Journal of Clinical Urology

摘　　要：目的:探讨多中心灶肾透明细胞癌各癌灶间的克隆起源关系。方法:提取20例多中心灶性肾透明细胞癌患者的共计42枚癌灶及对应正常组织的DNA后,10个微卫星多态性标志物D3s1038、D3s1234、D3s1300、D3s1317、D3s1540、D3s1597、D7s522、D8s261、D9s171、TP53作为引物对提取的DNA进行PCR扩增。用Labworks 3.0凝胶分析软件对PCR产物进行分析。在某一位点的信息性病例中若肿瘤组织的某一条等位基因条带消失或相对密度减少>50%,则记录为杂合性缺失(LOH)。观察各癌灶DNA的LOH类型。多中心病灶与原发灶显示相同的等位基因图谱示为共同克隆来源,否则为独立克隆起源。结果:在20例多中心灶性肾透明细胞癌中,17例(85%)在上述10位点中的至少1个位点检测到LOH,其中14例(14/17)检测到原发灶与相应的多中心灶有相同的LOH类型。在该14例中,有3例仅多中心癌灶在某一位点检测到LOH,另外有2例仅原发癌灶在某一位点检测到LOH。有3例(3/17)检测到原发灶与相应并发癌灶有不相同的LOH类型。结论:实验研究结果表明似乎大多数多中心灶性肾透明细胞癌为同一克隆起源,即由原发癌灶肾内转移而来。但是我们也发现了多克隆起源的证据。结合以往国内外相关研究,我们认为多中心灶性肾透明细胞癌同时存在单克隆起源和多克隆起源两种发生机制。Objective：To determine genetic relationships among the multifocal clear cell renal cell tumors by molecular genetic analysis. Method： A total of 42 tumors from 20 patients with multifocal clear cell renal cell carcinomas were investigated. All patients had multiple separate clear cell renal carcinomas. Loss of heterogeneity （LOH） analyses were done by PCR using ten markers for chromosome 3p25 （D3s1317, D3s1038, D3s1597）, 3p14 （D3s1300, D3s1234, D3s1540）, 7q31（D7s522）, 8q21 （D8s261）, 9q21 （D9s171）, and 17p13 （TP53）. PCR was performed according to standard protocols, followed by gel electrophoresis. The band patterns were observed and analyzed using Labworks 3.0. LOH was defined as a 〉50% reduction of the intensity in either of the two alleles as compared with those in normal control panels. PCRs for each polymorphic microsatellite marker were repeated at least twice from the same DNA preparations and the same results were obtained. Result： 17 of 20 （85%） patients with multifocal clear cell renal cell carcinoma showed allele loss in at least 1 of 10 microsatellite loci in separate tumors analyzed. Primary and satellite tumors showed a concordant allelic loss pattern in at least 1 informative locus in 14 of the 17 cases. In 3 of the 14 cases, LOH was observed only in satellite tumor lesions, and in 2 cases, LOH was detected only in primary lesions. A discordant pattern of allelic loss between coexisting kidney tumors was observed in three cases. Conclusion： Our data suggest that the majority of multifocal clear cell renal cell carcinomas have a common clonal origin. But we have also detected the evidence for polyclonal origin of multifocal clear cell renal cell carcinoma as well. Consequently, the multifocal clear cell renal cell carcinomas may be classified into monoclonal and polyclonal origin types.

关 键 词：肾透明细胞癌 多中心灶 LOH 克隆起源 

分 类 号：R737.11[医药卫生—肿瘤]