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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 investigate the origin of the clonal relationship between various cancerous lesions of multi-center renal clear cell carcinoma. Methods: Twenty microsatellite polymorphism markers, including D3s1038, D3s1234, D3s1300, D3s1317, D3s1540, D3s1597 and D7s522, were obtained from a total of 42 cancerous lesions and corresponding normal tissues in 20 patients with multicellular focal renal clear cell carcinoma , D8s261, D9s171, TP53 as primers for DNA amplification. PCR products were analyzed using Labworks 3.0 gel analysis software. Loss of heterozygosity (LOH) is recorded as informative cases at one site if one of the alleles of the tumor tissue disappears or the relative density decreases> 50%. Observe the DNA of each tumor LOH type. Multi-center lesions show the same allelic pattern as the primary tumor as a common clonal source, otherwise as an independent clonal origin. Results: In 20 cases of multicentric renal clear cell carcinoma, 17 cases (85%) detected LOH in at least one of the above 10 sites, of which 14 cases (14/17) detected primary tumors Same type of LOH as the corresponding multifocal heart. Of the 14 cases, LOH was detected in only one of the three cases in the multicentric foci, and only two in the primary foci. In 3 cases (3/17), different types of LOH were detected in the primary and corresponding concurrent foci. CONCLUSIONS: The experimental results show that it appears that most of the multicellular focal, clear-cell renal cell carcinomas are of the same clonal origin, that is, metastases from the primary foci within the kidney. However, we also found evidence of polyclonal origin. Combined with previous studies at home and abroad, we believe that multiple focal renal clear cell carcinoma exist both monoclonal origin and polyclonal origin mechanism.