GENETIC EVENTS IN HUMAN PROSTATE CELL TRANSFORMATION

描述(申请人描述)：本建议书的目标是 A)为以下人员提供系统的分子遗传学培训计划 申请人须发展为独立研究人员，以及b)研究 与克服衰老有关的遗传事件(即永生) 在人类前列腺上皮细胞中。大量的文献研究表明 前列腺癌基因组丢失和获得的详细模式。然而， 许多这些遗传事件或组合的功能意义 关于遗传事件的原因尚不清楚。克服衰老可以被认为是 肿瘤发生过程中有明确遗传基础的关键事件。在 建议的模型，人前列腺上皮细胞(HPECs)表达病毒 癌蛋白缺乏正常的pRb和/或p53调节功能(基因通常 在临床前列腺癌中发生改变)，并且寿命延长。这些 在体外实验中，这些细胞处于更高的自发性风险中 与永生有关的遗传和表观遗传改变。这个 申请者将测试这一假设，即这些基因的组合 补充P53和/或pRb丢失的事件在克服 体外前列腺癌细胞衰老的研究。此外，他们还建议 这些通路在前列腺癌的临床标本中被发现。他们的 具体目标包括：i)建立和表征体外模型 具有永生HPEC的系统，这些HPEC在功能上丢失了P53和/或Rb 选择性HPV16E6和/或E7逆转录病毒感染，II)识别 其他遗传和表观遗传事件，主要是事件的组合， 与克服P53和Rb连锁途径的衰老有关，III) 为了重新表达这些在永生化过程中失去或获得的基因组区域 永生和正常的HPECs，以及iv)将这些体外事件与 丢失P53和/或Rb的临床前列腺癌样本 功能。该提案将为研究分子遗传学提供新的视角。 以及通过P53和/或与克服衰老相关的表观遗传事件 在体外培养的前列腺上皮细胞中，Rb功能丧失。除了……之外 为了解决这一在人类前列腺癌中的关键机制作用，它 将为申请者提供结构化的分子技术培训计划 凯瑟琳·雷兹尼科夫博士的实验室里的遗传学。

DESCRIPTION (Applicant's Description): The objectives of this proposal are to a) provide a structured, training program in molecular genetics for the applicant to develop into an independent researcher, and b) study the genetic events associated with overcoming senescence (i.e. immortalization) in human prostate epithelial cells. Numerous documentational studies have detailed patterns of genomic loss and gain in prostate cancers. However, the functional significance of many of these genetic events or combinations of genetic events remains unclear. Overcoming senescence can be considered a critical event in tumorigenesis that has a clear genetic basis. In the proposed model, human prostate epithelial cultures (HPECs) expressing viral oncoproteins lack normal pRB and/or p53 regulatory functions (genes commonly altered in clinical prostate cancers) and have an extended lifespan. These in vitro events place these cells at high risk for additional 'spontaneous' genetic and epigenetic alterations associated with immortalization. The applicants will test the hypothesis that combinations of these genetic events, complementing p53 and/or pRb loss, are important in overcoming cellular senescence in in vitro prostate cancer. In addition, they propose that these pathways are found in clinical prostate cancer specimens. Their SPECIFIC AIMS include: I) to establish and characterize an in vitro model system with immortal HPECs that have functionally lost p53 and/or Rb by selective HPV16 E6 and/or E7 retroviral infection, ii) to identify additional genetic and epigenetic events, chiefly combinations of events, associated with overcoming senescence for p53- and Rb-linked pathways, iii) to reexpress these genomic regions lost or gained at immortalization in immortal and normal HPECs, and iv) to correlate these in vitro events with clinical prostate cancer samples that have lost either p53 and/or Rb function. The proposal will provide new i n sight into molecular genetic and epigenetic events associated with overcoming senescence, via p53 and/or Rb loss of function, in prostate epithelial cells in vitro. In addition to addressing this critical mechanistic role in human prostate neoplasia, it will provide the applicant with a structured training program in molecular genetics in the laboratory of Dr. Catherine Reznikoff.