Hatina J, Huckenbeck W, Rieder H, Seifert H-H, Schulz W A
Forschungslabor der Urologischen Klinik, Universitätsklinikum, Düsseldorf, Germany.
Urologe A. 2008 Jun;47(6):724-34. doi: 10.1007/s00120-008-1687-4.
Tumour cell lines represent valuable preclinical models to decipher underlying biology and identify potential therapy targets and pharmacologically useful compounds. Approximately 50 human bladder cancer cell lines have been established to date, mainly from invasive and metastatic tumours. Two of these, namely T24 and 253J, were experimentally further developed into progression series. These models have provided important insights into later tumour progression events and metastatic dissemination. Only a few cell lines are available as models of non-invasive papillary bladder cancer and no progression series have yet been established.
During the course of establishing a doxorubicin-resistant variant cell line of the human papillary bladder carcinoma cell line BFTC-905, a unique cell colony was identified, apparently involving cells with divergent growth patterns. Subsequent subculturing yielded three daughter cell lines, BFTC-905-compact, BFTC-905-diffuse und BFTC-905-diffuse M. Their fundamental characterization included basic cell morphology, cell membrane expression of E-Cadherin, karyotype analysis, invasion and colony forming capacity in soft agar. The clonal origin of the newly established daughter cell lines was assessed by means of molecular genetic methods.
We could identify important differences in multiple transformation related traits among the cell lines of the BFTC-905 progression series. Both diffuse cell lines (BFTC-905-diffuse und BFTC-905-diffuse M) differed from the BFTC-905-compact cell line by growing in a less organized,"diffuse" manner, which involved colonies of cells exhibiting apparently normal cell-to-cell adhesion as well as individual cells outside of them. This diminution of the cell-to-cell adhesion was accompanied by a corresponding decrease of membranous E-Cadherin. The BFTC-905-diffuse M cell line displayed a dramatic increase in the overall chromosome number, resulting in a hypertetraploid karyotype. At the same time, this cell line, as the only one in the progression series, acquired the ability to grow independent of anchorage in soft agar. All three cell lines remained noninvasive. Allelic distribution of highly polymorphic DNA-markers in the cell lines of the BFTC-905 progression series provided unequivocal evidence of their common origin.
The newly established BFTC-905 progression series manifests two aspects of the early progression of non-invasive bladder carcinoma, not exhibited by any other progression series published so far, namely dynamic changes in the expression of E-Cadherin and a complex karyotypic evolution. It may thus contribute important insights into further understanding of the pathobiology of bladder cancer.
肿瘤细胞系是用于解析潜在生物学机制、识别潜在治疗靶点和药理活性化合物的重要临床前模型。迄今为止，已建立了约50种人膀胱癌细胞系，主要来源于侵袭性和转移性肿瘤。其中两种，即T24和253J，经实验进一步发展成了进展系列。这些模型为深入了解肿瘤后期进展事件和转移扩散提供了重要见解。目前仅有少数细胞系可作为非侵袭性乳头状膀胱癌的模型，且尚未建立进展系列。
在建立人乳头状膀胱癌细胞系BFTC - 905的阿霉素耐药变异细胞系过程中，鉴定出一个独特的细胞集落，其细胞生长模式明显不同。随后传代培养得到三个子代细胞系，即BFTC - 905 - compact、BFTC - 905 - diffuse和BFTC - 905 - diffuse M。其基本特征包括基本细胞形态、E - 钙黏蛋白的细胞膜表达、核型分析、侵袭能力以及在软琼脂中的集落形成能力。通过分子遗传学方法评估新建立的子代细胞系的克隆起源。
我们发现BFTC - 905进展系列细胞系在多个与转化相关的特征上存在重要差异。两种弥漫性细胞系（BFTC - 905 - diffuse和BFTC - 905 - diffuse M）与BFTC - 905 - compact细胞系不同，它们以一种组织性较差的“弥漫性”方式生长，形成的细胞集落中细胞间黏附正常，集落外还有单个细胞。细胞间黏附的减弱伴随着细胞膜E - 钙黏蛋白的相应减少。BFTC - 905 - diffuse M细胞系的总染色体数显著增加，导致超四倍体核型。同时，该细胞系是进展系列中唯一获得在软琼脂中不依赖锚定生长能力的细胞系。所有三个细胞系均保持非侵袭性。BFTC - 905进展系列细胞系中高度多态性DNA标记的等位基因分布明确证明了它们的共同起源。
新建立的BFTC - 905进展系列体现了非侵袭性膀胱癌早期进展的两个方面，这是迄今已发表的任何其他进展系列均未展现的，即E - 钙黏蛋白表达的动态变化和复杂的核型进化。因此，它可能为进一步理解膀胱癌的病理生物学提供重要见解。
