DEFINING THE CELLULAR AND PHYSIOLOGICAL ROLE OF ABCA2

描述(申请人提供)：三磷酸腺苷结合盒(ABC)转运体ABCA2的过度表达与雌激素耐药有关，雌激素是一种用于治疗激素难治性前列腺癌的药物。ABCA2在将胆固醇、类固醇和脂类转运到内溶酶体中起着推定的作用，这表明它在包括癌症和动脉粥样硬化在内的人类疾病中发挥了作用。到目前为止，ABCA2在细胞系中的强制过表达一直是用于研究其功能的唯一工具。这项建议中概述的研究使用ABCA2基因敲除和敲入转基因小鼠来检验ABCA2在与前列腺癌TRAMP小鼠模型杂交的小鼠的胆固醇代谢、生理和前列腺癌发展/治疗反应中的作用。来自基因敲除和敲除小鼠的小鼠胚胎成纤维细胞(MEF)和稳定转染可诱导ABCA2表达或RNA干扰的前列腺癌细胞(PCS、DU145、LNCaP)将被分析对ABCA2水平变化的反应。ABCA2的表达对细胞增殖、胆固醇转运和药物治疗反应的影响将被研究。最终目标是利用转基因动物模型、MEF和人前列腺癌细胞来了解ABCA2的细胞和生理功能。

DESCRIPTION (provided by applicant): Overexpression of the ATP binding cassette (ABC) transporter, ABCA2, has been linked to resistance to estramustine, a drug used to treat hormone-refractory prostate cancer. ABCA2 has a putative role in trafficking cholesterol, steroids and lipids into endolysosomes, suggesting a role in human disease, including cancer and atherosclerosis. To date, forced over-expression of ABCA2 in cell lines has been the only tool used to investigate its function. The studies outlined in this proposal use ABCA2-knockout and -knock-in transgenic mice to examine the role of ABCA2 in cholesterol metabolism, physiology and prostate tumor development/treatment response in mice crossed to the TRAMP mouse model of prostate carcinoma. Mouse embryonic fibroblasts (MEFs), derived from knockout and knock-in mice, and prostate cancer cells (PCS, DU145, LNCaP), stably transfected with inducible ABCA2 expression or RNA interference, will be analyzed for response to changes in ABCA2 levels. Effects of ABCA2 expression on proliferation, cholesterol trafficking and drug treatment response will be examined. The ultimate goal is to understand the cellular and physiological function of ABCA2 using transgenic animal models, MEFs and human prostate cancer cells.