Williams Karin, Fernandez Suzanne, Stien Xavier, Ishii Kenichiro, Love Harold D, Lau Yun-Fai Chris, Roberts Richard L, Hayward Simon W
Department of Urologic Surgery, Vanderbilt University Medical Center, Nashville, Tennessee 37212-2765, USA.
Prostate. 2005 Jun 1;63(4):369-84. doi: 10.1002/pros.20200.
We have sought to develop a new in vivo model of prostate carcinogenesis using human prostatic epithelial cell cultures. Human prostate cancers frequently display DNA amplification in the 8q24 amplicon, which leads to an increase in the copy number of the c-MYC gene, a finding that suggests a role for c-MYC in human prostate carcinogenesis. In addition overexpression of c-MYC in transgenic mouse models results in prostatic carcinogenesis.
We took advantage of the ability of retroviruses to integrate foreign DNA into human prostatic epithelium (huPrE) to generate cell lines that overexpress the c-MYC protooncogene. These cells were recombined with inductive rat urogenital sinus mesenchyme and grafted beneath the renal capsule of immunocompromised rodent hosts.
The resultant tissue displayed a phenotype consistent with a poorly differentiated human prostatic adenocarcinoma. The tumors were rapidly growing with a high proliferative index. The neoplastic cells in the tumor expressed both androgen receptors (AR) and prostate-specific antigen (PSA), both characteristic markers of human prostate cancers. Microarray analysis of human prostatic epithelial cells overexpression c-MYC identified a large number of differentially expressed genes some of which have been suggested to characterize a subset of human cancers that have myc overexpression. Specific examples were confirmed by Western blot analysis and include upregulation of c-Myb and decreased expression of PTEN. Control grafts using either uninfected huPrE or using huPrE cells infected using an empty vector expressing a green fluorescent protein tag gave rise to well differentiated benign prostatic glandular ducts.
By using a retroviral infection strategy followed by tissue recombination we have created a model of human prostate cancer that demonstrates that the c-MYC gene is sufficient to induce carcinogenesis.
我们试图利用人前列腺上皮细胞培养物开发一种新的前列腺癌发生的体内模型。人类前列腺癌经常在8q24扩增子中显示DNA扩增，这导致c-MYC基因拷贝数增加，这一发现表明c-MYC在人类前列腺癌发生中起作用。此外，在转基因小鼠模型中c-MYC的过表达会导致前列腺癌发生。
我们利用逆转录病毒将外源DNA整合到人类前列腺上皮（huPrE）中的能力来生成过表达c-MYC原癌基因的细胞系。这些细胞与诱导性大鼠泌尿生殖窦间充质重组，并移植到免疫受损啮齿动物宿主的肾包膜下。
所得组织表现出与低分化人类前列腺腺癌一致的表型。肿瘤生长迅速，增殖指数高。肿瘤中的肿瘤细胞表达雄激素受体（AR）和前列腺特异性抗原（PSA），这两种都是人类前列腺癌的特征性标志物。对过表达c-MYC的人前列腺上皮细胞进行微阵列分析，鉴定出大量差异表达基因，其中一些已被认为可表征具有myc过表达的人类癌症子集。通过蛋白质免疫印迹分析证实了具体实例，包括c-Myb的上调和PTEN的表达降低。使用未感染的huPrE或使用用表达绿色荧光蛋白标签的空载体感染的huPrE细胞进行的对照移植产生了分化良好的良性前列腺腺管。
通过使用逆转录病毒感染策略然后进行组织重组，我们创建了一种人类前列腺癌模型，该模型表明c-MYC基因足以诱导癌发生。
