Basic study on the mechanism of intralaminal seeding of urothelial tumor using a short-term culture system for exfoliuted cells in patients urine

我们尝试培养尿脱落细胞，并阐明尿路上皮肿瘤细胞在腔内种植的机制。尿路上皮癌患者尿细胞原代培养成功率为87%，首次传代成功率为75%，明显高于健康志愿者的62%和35%。这种短期培养的成功率与肿瘤分级或分期无关。上尿路肿瘤患者的尿细胞原代培养成功率和首次传代成功率均为100%，膀胱肿瘤患者的尿细胞原代培养成功率和首次传代成功率均为65%（p<0.05）。尿细胞培养的成功率与肿瘤细胞中p53突变状态相关。p53阳性组的成功率为85%，p53阴性组的成功率为58%（P<0.05）。尿培养细胞p53状态的免疫细胞化学诊断结果与肿瘤细胞p53免疫组化染色结果高度一致（92.1%）。我们已成功地检测到相同的p53基因突变的尿液培养细胞来源的DNA中相应的肿瘤的突变等位基因特异性扩增。

We have tried to cultivate urinary exfoliated cells and to elucidate the mechanism of the intraluminal seeding of urothelial tumor cells. The sucess rates of both primary cell outgrowth and the first passage for urine cells from urothelial cancer patients were 87% and 75%, respectively, which were significantly higher than those for healthy volunteers (62% and 35%, respectively). The success rate of this short-term culture was not correlated with tumor grade or stage. Interestingly, the success rates for primary growth and the first passage were 100% for urine cells from the patients with upper urinary tract tumor, while those from bladder tumor patients were 65% (p<0.05). The success rates for urine cell culture were correlated with the p53 mutational status in tumor cells. The success rate for patients with p53 positive tumors was 85%, while that for patients with p53 negative tumors was 58% (p<0.05). The results of the immunocytochemical diagnosis for p53 status for urine cultured cells were highly identical (92.1%) with the results of the imunohistochemical staining of p53 for tumor cells in each patient. We have succeeded in detecting the same p53 gene mutation as the corresponding tumor in the urine culcure cell-derived DNA by the mutant allele-specific amplification.