P21 transcriptional inhibitors as proapoptotic compounds

描述（由申请人提供）： 放疗和化疗都是癌症治疗的重要方式。它 已经提出这些抗癌治疗通过以下方式消除肿瘤细胞： 诱导程序性细胞死亡（凋亡）。细胞周期蛋白依赖性激酶 抑制剂p21（Wafl/Cipl）在转录水平被诱导， 参与癌症发展的信号通路，并通过 各种抗癌治疗。越来越多的证据表明，p21是一种 细胞凋亡的一般抑制剂。我们假设抑制p21 肿瘤细胞中的转录将增强它们对辐射的反应， 因为p21的转录诱导通常使肿瘤 对这些治疗有抵抗力。我们提出了一个战略，以确定化学 p21启动子的抑制剂，我们计划测试有效的化学抑制剂， 肿瘤中p21转录与放化疗的关系 不同来源的细胞系和异种移植肿瘤，以评估其 通过p53依赖性和非依赖性凋亡对细胞杀伤的作用。细胞 与人p21启动子控制下的细菌LacZ基因一致， 其将通过p53依赖性机制被阿霉素高度诱导 将建立。该细胞系将用于鉴定化学物质 P21转录的抑制剂， 化学DIVERSet（TM）文库，其可从Chembridge化学获得 Corporation.抑制p21启动子的化合物将在本发明的方法中重新筛选。 含有lacZ报告基因的ConA细胞系在 ap 53-应答启动子，以确保它们不损害 p53作为转录激活因子。最有效的抑制剂 p21启动子的p53依赖性激活不影响p53依赖性 将在体外和体内鉴定和测试ConA细胞中LacZ的活化。 vivo.这些化合物将代表p21启动子的阻遏物，但不是 p53的抑制剂。这项研究的具体目的是确定化学品 p21转录的抑制剂，并将它们与 化疗药物在癌细胞系和异种移植肿瘤中的应用。我们预计 这些化合物将使肿瘤细胞对抗癌治疗敏感， 验证p21表达作为治疗靶点。这项研究可能会改善 通过鉴定新的化合物来治疗癌症， 将增加促进细胞死亡的抗癌药物的效率， 癌症治疗

DESCRIPTION (provided by applicant): Both radiation and chemotherapy are important modes of cancer treatment. It has been suggested that these anticancer treatments eliminate tumor cells by inducing programmed cell death (apoptosis). The cyclin-dependent kinase inhibitor p21 (Wafl/Cipl) is induced at the transcriptional level both by signaling pathways that participate in the development of cancer, and by a variety of anticancer treatments. There is a mounting evidence that p21 is a general inhibitor of apoptosis. We hypothesize that suppression of p21 transcription in tumor cells will enhance their response to radiation and chemotherapy because transcriptional induction of p21 usually makes tumors resistant to these treatments. We propose a strategy to identify chemical inhibitors of p21 promoter and we plan to test potent chemical inhibitors of p21 transcription together with radiation and chemotherapeutic drugs in tumor cell lines of different origin and in xenograft tumors to evaluate their effect on cell killing via p53-dependent and -independent apoptosis. A cell line with the bacterial LacZ gene under control of the human p21 promoter, which will be highly inducible by adriamycin via a p53-dependent mechanism will be established. This cell line will be used for identifying chemical inhibitors of p21 transcription by screening of individual compounds of a chemical DlVERSet(TM) library that is commerically available from Chembridge Corporation. Compounds that repress the p21 promoter will be rescreened in a ConA cell line containing the lacZ reporter gene under the control of ap53-responsive promoter to ensure that they do not compromise the ability of p53 to act as a transcriptional activator. The most potent inhibitors of p53-dependent activation of the p21 promoter that do not affect p53-dependent activation of LacZ in ConA cells will be identified and tested in vitro and in vivo. These compounds will represent repressors of the p21 promoter, but not inhibitors of p53. The specific aims of the study are to identify chemical inhibitors of p21 transcription and to test them in combination with chemotherapeutic drugs in cancer cell lines and xenograft tumors. We expect that such compounds will sensitize tumor cells to anti-cancer therapy, validating p21 expression as a therapeutic target. This study may improve treatment of cancer by leading to the identification of new compounds that will increase the efficiency of cell death promoting anticancer drugs for cancer treatment.