Chaffer Christine L, Dopheide Bonnie, McCulloch Daniel R, Lee Allan B, Moseley Jane M, Thompson Erik W, Williams Elizabeth D
Bernard O'Brien Institute of Microsurgery, University of Melbourne, Australia.
Clin Exp Metastasis. 2005;22(2):115-25. doi: 10.1007/s10585-005-5141-3.
Muscle invasive transitional cell carcinoma (TCC) of the bladder is associated with a high frequency of metastasis, resulting in poor prognosis for patients presenting with this disease. Models that capture and demonstrate step-wise enhancement of elements of the human metastatic cascade on a similar genetic background are useful research tools. We have utilized the transitional cell carcinoma cell line TSU-Pr1 to develop an in vivo experimental model of bladder TCC metastasis. TSU-Pr1 cells were inoculated into the left cardiac ventricle of SCID mice and the development of bone metastases was monitored using high resolution X-ray. Tumor tissue from a single bone lesion was excised and cultured in vitro to generate the TSU-Pr1-B1 subline. This cycle was repeated with the TSU-Pr1-B1 cells to generate the successive subline TSU-Pr1-B2. DNA profiling and karyotype analysis confirmed the genetic relationship of these three cell lines. In vitro, the growth rate of these cell lines was not significantly different. However, following intracardiac inoculation TSU-Pr1, TSU-Pr1-B1 and TSU-Pr1-B2 exhibited increasing metastatic potential with a concomitant decrease in time to the onset of radiologically detectable metastatic bone lesions. Significant elevations in the levels of mRNA expression of the matrix metalloproteases (MMPs) membrane type 1-MMP (MT1-MMP), MT2-MMP and MMP-9, and their inhibitor, tissue inhibitor of metalloprotease-2 (TIMP-2), across the progressively metastatic cell lines, were detected by quantitative PCR. Given the role of MT1-MMP and TIMP-2 in MMP-2 activation, and the upregulation of MMP-9, these data suggest an important role for matrix remodeling, particularly basement membrane, in this progression. The TSU-Pr1-B1/B2 model holds promise for further identification of important molecules.
膀胱肌层浸润性移行细胞癌（TCC）常伴有高频率转移，导致该疾病患者预后较差。在相似遗传背景下能够捕捉并展示人类转移级联反应各要素逐步增强的模型是有用的研究工具。我们利用移行细胞癌细胞系TSU-Pr1建立了膀胱TCC转移的体内实验模型。将TSU-Pr1细胞接种到SCID小鼠的左心室，使用高分辨率X射线监测骨转移的发生情况。切除单个骨病变处的肿瘤组织并在体外培养，以产生TSU-Pr1-B1亚系。用TSU-Pr1-B1细胞重复此过程，以产生后续亚系TSU-Pr1-B2。DNA谱分析和核型分析证实了这三种细胞系的遗传关系。在体外，这些细胞系的生长速率没有显著差异。然而，心内接种后，TSU-Pr1、TSU-Pr1-B1和TSU-Pr1-B2的转移潜能逐渐增加，同时出现可通过放射学检测到的转移性骨病变的时间相应缩短。通过定量PCR检测发现，在逐渐具有转移能力的细胞系中，基质金属蛋白酶（MMPs）膜型1-MMP（MT1-MMP）、MT2-MMP和MMP-9及其抑制剂金属蛋白酶组织抑制剂-2（TIMP-2）的mRNA表达水平显著升高。鉴于MT1-MMP和TIMP-2在MMP-2激活中的作用以及MMP-9的上调，这些数据表明基质重塑，尤其是基底膜重塑，在这一进展过程中起重要作用。TSU-Pr1-B1/B2模型有望进一步鉴定重要分子。
