Molecular biological analysis of urothelial cancers using short-term cultures of urinary exfoliated cells

尿路上皮癌，特别是上尿路癌，是异位尿路上皮的多灶性同步和/或异时性肿瘤。我们研究了尿路上皮癌患者尿液中脱落细胞在体外定植和增殖的能力。对129例尿路上皮癌患者和53例健康志愿者的尿液样本进行短期培养，比较了尿路上皮脱落细胞的生长潜力和临床病理特征，特别是肿瘤定位和腔内肿瘤复发的证据。来自患者的129个细胞培养中有112个（86.8%）出现原代细胞生长，来自健康志愿者的53个细胞培养中有29个（54.7%）出现原代细胞生长（p<0.0001）。来自患者的129个培养物中有77个（59.7%）获得了“足够的细胞生长”（每瓶超过10^5个细胞），来自健康志愿者的53个培养物中有7个（13.2%）获得了“足够的细胞生长”（p<0.0001）。在肿瘤定位方面，上尿路肿瘤患者的细胞生长率（21/25,84.0%）显著高于膀胱肿瘤患者（56/104,53.8%）（p=0.0062）。比例风险回归分析显示，只有膀胱脱落细胞的生长势能显著预测浅表性膀胱肿瘤患者膀胱内肿瘤复发（p=0.011）。尿路上皮癌的腔内多灶性肿瘤复发与尿路上皮脱落细胞定植和增殖的潜力有关。p53基因突变和17p染色体等位基因缺失与尿路上皮癌的高分级和侵袭性密切相关。采用短期培养系统对p53进行免疫细胞化学检测。92.1%患者尿细胞p53免疫化学分析结果与肿瘤细胞p53免疫化学分析结果一致。尿中脱落细胞培养是一种很好的尿路上皮癌分子诊断方法。特别是，它是一种非侵入性检查，对于p53突变肿瘤的早期发现和强化随访是有用的。少

Urothelial cancers, in particular of upper urinary tract cancers, is multifocal synchronous and/or metachronous tumor development in the heterotopic urothelium. We investigated the abilities of exfoliated cells in the urine of patients with urothelial cancers to colonize and proliferate in vitro. Short-term cultures of 129 urine samples obtained from patients with urothelial cancers and 53 samples from healthy volunteers were compared as to the growth potential of urinary-exfoliated cells and clinico-pathological features, in particular tumor localization and evidence of intraluminal tumor recurrence. Primary cell outgrowth occurred in 112 (86.8%) of the 129 cell cultures from the patients and in 29 (54.7%) of the 53 from the healthy volunteers (p<0.0001). "Sufficient cell growth" (more than 10^5 cells per flask) was obtained for 77 (59.7%) of the 129 cultures from patients and 7(13.2%) of the 53 from the healthy volunteers (p<0.0001). In terms of tumor localization, the rate of suffic … More ient cell growth was significantly higher for patients with upper urinary tract tumor (21/25,84.0%) than for those with bladder tumor (56/104,53.8%) (p=0.0062). Proportional hazard regression analysis showed that only the growth potential of the urinary-exfoliated cells was significantly predictive of intravesical tumor recurrence in patients with superficial bladder tumors (p=0.011). The potentials for the colonization and proliferation of urinary-exfoliated cells are associated with intraluminal multifocal tumorrecurrence of urothelial cancers.Mutation of the p53 gene together with allelic loss of chromosome 17p correlates well with high grade and invasiveness of urothelial cancer. Using the short-term culture system, immunocytochemical detection of p53 was conducted. Results of immunochemical analyzes of the p53 in the urine cells and those in the tumor samples were identical in 92.1% of the patients. Culture of exfoliated cells in urine is a good method for the molecular diagnosis of urothelial cancer. In particular, it is a non-invasive examination that should prove useful for the early detection and intensive follow-up of tumorswith p53 mutation. Less