CELLULAR AND ANIMAL STUDIES OF SRC INHIBITORS

该计划提供了假定的抑制剂的生物模型， 人结肠腺癌细胞系和由这些细胞衍生的肿瘤 线 我实验室过去的工作表明， pp 6c-src和pp 62 c-yes在人类结肠肿瘤中是一致的， pp 60 c-src对结肠癌细胞系的致瘤性具有重要作用， 裸鼠，以及充分表征的人结肠肿瘤细胞系， HT 29在筛选PTK抑制剂方面非常有用。 因此该 该计划将利用我们所描述的系统来筛选假定的 在第一和第二个计划中开发的PTK抑制剂。 这些屏幕 将包括测定化合物在正常 成纤维细胞以及结肠肿瘤细胞中，这些细胞的生长抑制 细胞，以及对软琼脂集落形成的影响。 而且这个 该提案将检查受抑制细胞中src激酶的活性， 确定抑制与预测的机制相对应。 该项目还将在M.D.进行致瘤性研究。 安德森癌症中心，后肢和结肠原位模型 裸鼠的癌症。 抑制剂对pp 60 c-src的特异性将是 通过检查假定的PTK抑制剂在罕见的 没有激活的pp 60 c-src的结肠肿瘤细胞系。 结合 我的实验室的基础研究工作，这个筛选将测试 pp 60 c-src特异性抑制剂将降低致瘤性假设 的模型结肠肿瘤细胞系在没有显着的毒性。 几种铅化合物目前在测试的各个阶段都很少见，所以每种 各种分析将同时进行。 随着对 在这个建议中的其他方案，我们应该确定的功效， 用于结肠癌治疗性治疗的SRC特异性抑制剂， 了解肿瘤的分子特征， 抑制剂可能是有效的，并进一步了解 src家族PTKs在结肠肿瘤进展中的作用

This Program provides the biologic models for putative inhibitors using human colon adenocarcinoma cell lines and tumors derived from these cell lines. Past work from my laboratory has demonstrated that activation of pp6c-src and pp62c-yes are consistent in human colon tumors, that activated pp60c-src is important to the tumorigenicity of colon tumor cell lines in nude mice, and that well characterized human colon tumor cell lines such as HT 29 are very useful in screening putative PTK inhibitors. Thus this Program will utilize the systems we have described for screening putative PTK inhibitors developed in the first and second Programs. These screens will include determination of cytotoxicity of the compounds in normal fibroblasts as well as in colon tumor cells, growth inhibition of these cells, and effects on soft agar colony formation. Additionally, this proposal will examine the src kinase activity in inhibited cells to determine that the inhibition corresponds with the predicted mechanism. The program will also perform the tumorigenicity studies at the M.D. Anderson Cancer Center, in hind limb and in orthotopic models for colon cancer in nude mice. Specificity of inhibitors toward pp60c-src will be assessed by examining the effects of the putative PTK inhibitors in rare colon tumor cell lines without activated pp60c-src. In combination with the basic research efforts of my laboratory, this screening will test the hypothesis that a pp60c-src-specific inhibitor will reduce tumorigenicity of model colon tumor cell lines in the absence of significant toxicity. Several lead compounds rare currently at various stages of testing, so each of the various analyses will proceed simultaneously. With the studies of the other Programs in this proposal, we should determine the efficacy of src-specific inhibitors for therapeutic treatment of colon cancer, provide an understanding of the molecular profile of tumors inn which such inhibitors are likely to be affective, and further an understanding of the roles of src family PTKs in colon tumor progression.