Profiling the evolution of human metastatic bladder cancer
Profiling the evolution of human metastatic bladder cancer
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DOI：
10.1158/0008-5472.can-04-0826
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发表时间：
2004-11-01
期刊：
影响因子：
11.2
通讯作者：
Theodorescu, D
中科院分区：
文献类型：
--
作者：
Nicholson, BE;Frierson, HF;Theodorescu, D
侵袭性膀胱癌患者经常发生肺转移，然而在分子水平上对这一过程的研究受到人类转移肿瘤组织缺乏可用性和缺乏合适的动物模型的影响。为了解决这个问题，我们开发了越来越多的转移性人类膀胱癌细胞系和体内膀胱癌肺转移模型，并成功地利用这些模型鉴定了根据肺转移潜力程度而改变表达水平的基因。首先将低转移性的T24T人尿路上皮癌细胞静脉注射到裸鼠体内，然后从由此产生的小鼠肺肿瘤中连续重新引入和再分离人肿瘤细胞，依次分离出三种转移性越来越强的人类衍生细胞系，分别为FL1、FL2和FL3。为了鉴定与大多数肺转移表型相关的基因，来自亲本和衍生细胞的RNA补体用寡核苷酸微阵列进行了评估。在这样做的过程中，我们发现121个基因在T24T到FL3的过渡过程中逐渐上调，而43个基因逐渐下调。正如预期的那样，在这些群体中发现的许多基因，根据其蛋白质产物的归属功能，理论上可以参与组织侵袭和转移。此外，在转移转移过程中观察到的基因表达变化幅度与体内早期肺定植倾向和降低宿主存活率相关。为了进一步确定实验系统中发现的哪些基因与人膀胱癌肺转移相关，我们评估了23例不同分期和分级的原发性人膀胱癌的基因表达谱，然后将这些基因表达谱与模型细胞系中改变的基因表达谱进行了比较。在这里，我们发现表调节蛋白、尿激酶型纤溶酶原激活物（uPA）、基质金属蛋白酶（MMP）14和金属蛋白酶组织抑制剂（TIMP-2）的表达在患者组织中作为肿瘤分期的功能，与小鼠肺模型中观察到的转移潜力相比，一致且逐渐上调。这四种标志物在实验和临床情况之间的强相关性有助于验证该系统作为研究肺转移的有用工具，并确定治疗靶点，可能会减少患者这一过程的发生率。
Pulmonary metastases frequently develop in patients with aggressive bladder cancer, yet investigation of this process at the molecular level suffers from the poor availability of human metastatic tumor tissue and the absence of suitable animal models. To address this, we developed progressively more metastatic human bladder cancer cell lines and an in vivo bladder-cancer lung-metastasis model, and we successfully used these to identify genes of which the expression levels change according to the degree of pulmonary metastatic potential. By initially intravenously injecting the poorly metastatic T24T human urothelial cancer cells into nude mice, and then serially reintroducing and reisolating the human tumor cells from the resultant mouse lung tumors, three derivative human lines with increasingly metastatic phenotypes, designated FL1, FL2, and FL3, were sequentially isolated. To identify the genes associated with the most lung-metastatic phenotype, the RNA complement from the parental and derivative cells was evaluated with oligonucleotide microarrays. In doing so, we found 121 genes to be progressively up-regulated during the transition front T24T to FL3, whereas 43 genes were progressively down-regulated. As expected, many of the genes identified in these groups could, according to the ascribed functions of their protein product, theoretically participate in tissue invasion and metastasis. In addition, the magnitude of gene expression changes observed during the metastatic transition correlated with the in vivo propensity for earlier lung colonization and decreased host survival. To additionally define which genes found in the experimental system were of relevance to human bladder cancer lung metastasis, we evaluated gene expression profiles of 23 primary human bladder tumors of various stages and grades, and then we compared these gene expression profiles to the altered profiles in our model cell lines. Here we found that the expression of epiregulin, urokinase-type plasminogen activator (uPA), matrix metalloproteinase (MMP)14, and tissue inhibitor of metalloproteinase (TIMP-2) were consistently and progressively up-regulated when viewed as a function of tumor stage in tissues of patients versus the metastatic potential seen in the mouse lung model. The strong correlation of these four markers between the experimental and clinical situations helps validate this system as a useful tool for the study of lung metastasis and defines targets of therapy that may reduce the incidence of this process in patients.