Berthon P, Cussenot O, Hopwood L, Leduc A, Maitland N
HOP ST LOUIS,DEPT UROL,F-75475 PARIS 10,FRANCE.
Int J Oncol. 1995 Feb;6(2):333-43. doi: 10.3892/ijo.6.2.333.
To study mesenchymal-epithelial interactions associated with the normal and pathological human prostate, we have developed a model of well differentiated human prostate epithelial and fibroblastic cells. Normal human prostatic cells, either of epithelial or fibroblastic origins were successfully transfected with SV40 and strains with extended lifespan were selected until the crisis was reached, within 20 and 30 passages for the epithelial and fibroblastic cells, respectively. Only a few clones emerged from the crisis: PNT1A (Cussenot et al: J Urol 143: 881-886, 1991), PNT1B and PNT2 epithelial cell lines. Successful immortalisation was achieved only with SV40 expressing both large T and small t oncogenes, while attempts to immortalise with a vector expressing SV40 large T alone have given a few strains showing no extended lifespan and no cells which overcame the crisis. A PNT2 subclone named PNT2-LSD which developed spontaneously (less serum dependent) was selected, characterised and included in the analysed series. The epithelial cell lines displayed a differentiation pattern which has been classified as follows (from high to low): PNT2>PNT2-LSD>PNT1A>PNT1B. Differentiation features studied were (i) the colony-forming ability of the PNT2 and PNT2-LSD compared to PNT1A and PNT1B, (ii) their respective doubling time of 39, 29, 30 and 28 hours, (iii) their decreasing serum dependency, (iv) the expression of cytokeratin 19 (a feature of well differentiated luminal cells of the glandular prostate) for PNT2 and PNT2-LSD. Furthermore, the mesenchymal derived pflsv1 cells were confirmed to be of fibroblastic nature. None of the cell lines analysed showed any tumourigenicity in nude mice over a period of 12 months. Serum deprivation and direct steroid withdrawal during the culture triggered cell death by apoptosis, an event which could be overcome by EGF stimulation, particularly for the well differentiated PNT2 cells. This interesting characteristic, which is similar to the high apoptotic rate observed ipl vivo for normal prostate, particularly after castration should lead, together with the other properties of these cell lines, to a better understanding of the biology of the different cell compartments involved in the progression of prostate towards neoplasia.
为了研究与正常和病理状态下的人类前列腺相关的间充质 - 上皮相互作用，我们建立了一个高分化人类前列腺上皮细胞和成纤维细胞的模型。将正常人类前列腺细胞（上皮来源或成纤维来源）用SV40成功转染，并分别在上皮细胞传代20次、成纤维细胞传代30次时选择寿命延长的菌株，直至达到危机期。危机期后仅出现了少数克隆：PNT1A（Cussenot等人：《泌尿学杂志》143：881 - 886，1991）、PNT1B和PNT2上皮细胞系。只有同时表达大T和小t癌基因的SV40才能成功实现永生化，而用仅表达SV40大T的载体进行永生化的尝试仅得到了少数寿命未延长且未克服危机期的菌株。我们选择了一个自发产生的（血清依赖性较低）PNT2亚克隆，命名为PNT2 - LSD，对其进行了表征并纳入分析序列。上皮细胞系表现出的分化模式如下（从高到低）：PNT2 > PNT2 - LSD > PNT1A > PNT1B。所研究的分化特征包括：（i）与PNT1A和PNT1B相比，PNT2和PNT2 - LSD的集落形成能力；（ii）它们各自的倍增时间分别为39、29、30和28小时；（iii）它们对血清依赖性的降低；（iv）PNT2和PNT2 - LSD中细胞角蛋白19的表达（这是腺性前列腺高分化腔面细胞的一个特征）。此外，间充质来源的pflsv1细胞被证实具有成纤维细胞性质。在12个月的时间里，所分析的细胞系在裸鼠中均未显示出任何致瘤性。培养过程中的血清剥夺和直接撤去类固醇会引发细胞凋亡导致细胞死亡，而这一过程可通过表皮生长因子（EGF）刺激来克服，尤其是对于高分化的PNT2细胞。这一有趣的特征，类似于在体内观察到的正常前列腺尤其是去势后的高凋亡率，与这些细胞系的其他特性一起，将有助于更好地理解前列腺向肿瘤发展过程中不同细胞成分的生物学特性。
