Webber M M, Bello D, Kleinman H K, Wartinger D D, Williams D E, Rhim J S
Department of Medicine, Michigan State University, East Lansing, USA.
Carcinogenesis. 1996 Aug;17(8):1641-6. doi: 10.1093/carcin/17.8.1641.
Progress in prostate cancer research has been hindered by the lack of well characterized, immortalized, human prostatic epithelial cell lines that express markers of normal prostatic epithelial cells and mimic normal growth and differentiation responses to androgens. The objectives of this study were to: (i) establish immortalized cell lines from non-neoplastic, adult human prostatic epithelium using adenovirus-12/simian virus-40 (Ad12-SV40) hybrid virus; (ii) establish their prostatic epithelial origin; (iii) demonstrate androgen responsiveness; and (iv) examine response to growth factors. Primary epithelial cell cultures derived from a non-neoplastic, adult human prostate were infected with the Ad12-SV40 virus. Several immortalized clones were isolated. Single cell cloning of one clone, free of cytopathic effects, gave rise to the PWr-1E cell line. An immortalized cell line PWR-1E, which expresses many characteristics of normal prostatic epithelial cells was established. Immunostaining showed that cells express cytokeratins 8 and 18 normally expressed by differentiated, secretory prostatic epithelial cells. The most remarkable characteristics of PWR-1E cells are growth stimulation, increased expression of androgen receptor and induction of prostate specific antigen (PSA) expression in response to androgens, which indisputably establish their prostatic epithelial origin. They are positive for SV40 large-T antigen and show strong nuclear staining for p53. Cells from passages 23 and 40 were not tumorigenic in nude mice even when co-injected with Matrigel. They grow in a serum-free defined medium and respond to EGF, bFGF and TGF-beta. Passage 42-cells showed a human male (XY), hyperdiploid karyotype. The PWR-1E cell line is the only known Ad12-SV40-immortalized human prostatic epithelial cell line. PWR-1E cells can be used to study (i) the etiology and the multistep process of carcinogenesis and tumor progression in the human prostate; (ii) normal prostate physiology and differentiation; and (iii) potential prostate cancer chemopreventive agents.
前列腺癌研究进展一直受到阻碍，因为缺乏特征明确、永生化的人前列腺上皮细胞系，这些细胞系要表达正常前列腺上皮细胞的标志物，并模拟对雄激素的正常生长和分化反应。本研究的目的是：（i）使用腺病毒12/猴病毒40（Ad12-SV40）杂交病毒从非肿瘤性成人前列腺上皮建立永生化细胞系；（ii）确定其前列腺上皮起源；（iii）证明雄激素反应性；（iv）检测对生长因子的反应。用Ad12-SV40病毒感染从非肿瘤性成人前列腺获得的原代上皮细胞培养物。分离出几个永生化克隆。对一个无细胞病变效应的克隆进行单细胞克隆，得到了PWr-1E细胞系。建立了一种表达许多正常前列腺上皮细胞特征的永生化细胞系PWR-1E。免疫染色显示细胞表达分化的分泌性前列腺上皮细胞正常表达的细胞角蛋白8和18。PWR-1E细胞最显著的特征是生长受刺激、雄激素受体表达增加以及对雄激素反应时前列腺特异性抗原（PSA）表达的诱导，这无可争议地确定了它们的前列腺上皮起源。它们对SV40大T抗原呈阳性，对p53呈强核染色。第23代和第40代细胞即使与基质胶共同注射也不会在裸鼠中致瘤。它们在无血清限定培养基中生长，并对表皮生长因子（EGF）、碱性成纤维细胞生长因子（bFGF）和转化生长因子-β（TGF-β）有反应。第42代细胞显示为人类男性（XY）超二倍体核型。PWR-1E细胞系是唯一已知的Ad12-SV40永生化人前列腺上皮细胞系。PWR-1E细胞可用于研究：（i）人类前列腺癌发生和肿瘤进展的病因及多步骤过程；（ii）正常前列腺生理学和分化；（iii）潜在的前列腺癌化学预防剂。
