The Role of AP-1 and Other Transcription Factors in Canc

这项研究的总体目标是识别和描述在肿瘤促进和肿瘤进展过程中推动限速步骤的基因调控事件。AP-1转录因子是Jun和Fos家族蛋白的异源二聚体，与某些基因转录启动子上的特定序列结合并驱动其转录。我们对小鼠JB6细胞和工程小鼠模型(Bernstein和Colburn，Science，1989，Dong等人PNAS，1994，Young等人，PNAS，1999)的观察证实，AP-1的激活是皮肤肿瘤发生和肿瘤进展所必需的。转基因小鼠角质形成细胞特异性表达显性阴性Jun抑制AP-1诱导的肿瘤形成，而不抑制与人类致癌相关的多种小鼠模型的生长、分化或增殖。其中包括因人类乳头瘤病毒E7的表达而提高皮肤肿瘤促进反应的小鼠(Young等人Molec Carc 2002)和通过反复暴露于UVB而诱导形成鳞癌的小鼠(Cooper等人Molec癌症RES 2003)。为了进一步研究，在合作者鲍威尔·布朗和杰伊·蒂施拉尔的实验室里，四环素调控的67的表达正被定向到乳腺和肺上皮细胞。当转基因K14-TAM67或四环素调控的TAM67结构在更先进的致瘤或锚定非依赖且显示AP-1和NFkB活性升高的人类细胞系中表达时，肿瘤细胞的表型被抑制(Li等人，Oncogene，1998和Li等人，Molec Cancog2000)。转录因子NFkappa B与AP-1协调调节，表明这两种因子在转化中可能具有重要作用(Li et，Cancer res 1997)。最近的观察发现，NFkB无反应性是JB6模型中转化无反应性的一个解释(Hsu等人，癌症RES 2001，Hu等人癌症发生2004年)。转化抵抗细胞的转化无反应归因于不能激活NFkappa B p65蛋白。S536处的p65磷酸化对于DNA结合和泛素化以及抑制物IkappaBα的降解都是重要的(Hu等人Molec癌症2005年)。因此，以AP-1和NFkB的上调为靶点阻止肿瘤的促进和进展的观察已经从小鼠JB6模型扩展到小鼠和人类角质形成细胞进展模型，以及转基因小鼠模型。对关键分子相互作用的新理解正在出现。表达AP-1/NFkB抑制因子67的转基因小鼠为鉴定AP-1或NFkB在肿瘤转化中起关键作用的靶基因提供了一个有价值的机会。表达芯片分析揭示了潜在的TAM67靶基因，这些基因是目前研究的对象。这样的靶基因可能是癌症预防的新的分子靶点(Young等人在2003年的Molec医学趋势中)。最近的研究排除了iNOS(Dhar等人癌症治疗2003年)，其他研究已经确定染色质结构蛋白HMGA1(Dhar等人癌基因2004)作为重要的TAM67靶标的重要性。针对确定激活AP-1所需的MAP激酶ERK依赖的分子事件的研究已经确定Fos家族蛋白Fra-1的激活是一个关键事件(Young等人，Molec Cell Biol 2002)。缺乏Erk或Fra-1蛋白的JB6变异体为AP-1和转化无反应。ERK和Fra-1的表达分别恢复了细胞的反应性。AP-1依赖基因启动子的一个子集预计需要Fra-1进行转录激活，这些启动子可能成为癌症预防的特别有效的靶点。

The overall aim of this research is to identify and characterize gene regulation events that propel rate-limiting steps during tumor promotion and tumor progression. The AP-1 transcription factor is a heterodimer of Jun and Fos family proteins that binds to a specific sequence on the transcriptional promoter of certain genes and drives their transcription. Our observations with mouse JB6 cells and with an engineered mouse model (Bernstein and Colburn, Science, 1989, Dong et al PNAS, 1994, Young et al., PNAS, 1999) established that AP-1 activation is required for skin tumorigenesis and tumor progression. Keratinocyte-specific expression of Dominant Negative Jun in transgenic mice inhibits induced AP-1 and tumorigenesis without inhibiting growth, differentiation or hyperplasia in multiple mouse models relevant to human carcinogenesis. Among these are mice whose skin tumor promotion response is elevated by expression of Human Papilloma Virus E7 (Young et al Molec Carc 2002) and mice induced to form squamous carcinomas by repeated exposure to UVB (Cooper et al Molec Cancer Res 2003). For further studies, tetracycline regulated expression of TAM 67 is being directed to mammary and lung epithelia in the laboratories of collaborators Powel Brown and Jay Tischlaar. When the transgene K14-TAM67 or a tetracycline regulated TAM67 construct was expressed in the more progressed human cell lines that are tumorigenic or anchorage independent and show elevated AP-1 and NFkB activities, tumor cell phenotype was suppressed (Li et al., Oncogene, 1998 and Li et al., Molec Carcinog 2000). The transcription factor NFkappa B is coordinately regulated with AP-1, suggesting the possible importance of both factors in transformation (Li et, Cancer Res 1997). Recent observations have identified NFkB non-responsiveness as an explanation for transformation non-responsiveness in the JB6 model (Hsu et al, Cancer Res 2001, Hu et al Carcinogenesis 2004). Transformation resistant cells owe their transformation nonresponsiveness to an inability to activate NFkappa B p65 protein. p65 phosphorylation at S536 is important both for DNA binding and for ubiquitination and degradation of inhibitor IkappaB alpha (Hu et al Molec Carcinog 2005). Thus the observation that targeting AP-1 and NFkB elevation prevents tumor promotion and progression has been extended from the mouse JB6 model to mouse and human keratinocyte progression models, and to transgenic mouse models. New understanding of critical molecular interactions is emerging. Transgenic mice expressing AP-1/ NFkB inhibitor TAM 67 present a valuable opportunity to identify AP-1 or NFkB target genes whose expression is critical to neoplastic transformation. Expression microarray analysis has revealed potential TAM67 target genes that are subjects of current research. Such target genes may be promising new molecular targets for cancer prevention (Young et al Trends in Molec Medicine 2003). Recent studies have excluded iNOS (Dhar et al Mol Cancer Ther 2003)and other studies have established the importance of chromatin architectural protein HMGA1(Dhar et al Oncogene 2004) as a functionally significant TAM67 target. Research directed to identifying MAP kinase ERK dependent molecular events required for activating AP-1 has identified the activation of Fos family protein Fra-1 as a pivotal event (Young et al, Molec Cell Biol 2002). JB6 variants deficient in either Erk or Fra-1 proteins are AP-1 and transformation nonresponsive. Responsiveness is restored by expression of Erk or Fra-1 respectively. A subset of AP-1 dependent gene promoters is expected to require Fra-1 for transcriptional activation and these may serve as particularly effective targets for cancer prevention.