ONCOGENIC ROLE OF JUN KINASE IN HUMAN PROSTATE CANCER

Jun 激酶在人类前列腺癌中的致癌作用

• 批准号: 6194816
• 负责人: DAN MERCOLA
• 金额: $ 37.56万
• 依托单位: SIDNEY KIMMEL CANCER CENTER
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-07-31 至 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6194816
• 关键词: JUN kinase; androgens; antisense nucleic acid; cell line; cell proliferation; cis platinum compound; complementary DNA; gene expression; genetic library; genetically modified animals; human tissue; immunocytochemistry; laboratory mouse; male; microarray technology; neoplastic transformation; oligonucleotides; oncoproteins; prostate neoplasms; protooncogene

The Jun Kinase/Stress-activated protein kinase pathway (JNK/SAPK) has been implicated as mediating stress-responses, survival, proliferation, and apoptosis. Previous studies, principally by C. Der and co-workers and M. Karin, in which this laboratory participated, have shown the JNK pathway is required for the transformation of primary fibroblasts by the activated ras oncogene and related oncogenes. We have extended the JNK analysis to human tumor cells. JNK is commonly serum-inducible or constitutively active in a variety of human tumor cells. Specific inhibition blocks tumor cell growth. Moreover, stable expression of a dominant negative inhibitor, c-Jun (S63A, S73A), blocks DNA repair and greatly sensitizes human tumor lines (T98G glioblastoma, A549 lung, MCF-7 breast, and PC3 prostate carcinoma) to killing by cisplatin, up to 9.7-fold for PC3 cells. We hypothesize that during progression of cancer, the JNK pathway is commonly selected as greatly enhancing DNA repair and synthesis, thereby facilitating oncongenesis. To test this, highly specific antisense compounds complementary to the two isoform families JNK1 and JNK2 have been developed and characterized. Systemic antisense treatment of athymic mice bearing established PC3 xenografts inhibits growth by 79 percent, better than cisplatin (49 percent), and combined antisense treatment promotes complete regression in high frequency. Conversely, PC3 cells are 100 percent tumorigenic. To test the generality, eight additional prostate cell lines were found to proliferate in direct proportion to the serum-inducible level of JNK activity over a 9-fold range. Moreover antisense JNK2 but not antisense JNK1 nearly completely blocks the growth of all lines in proportion to the JNK activity strongly indicating that JNK2 is commonly required for serum-stimulated growth prostate carcinoma cells. It is proposed to test the hypothesis that JNK is oncogenic in prostate carcinoma. We will test whether JNK is required for proliferation of prostate carcinoma lines in vitro (Aim 1) and in vivo (Aim 2). The efficacy of antisense JNK will be tested critically in the TRAMP model (Aim 2). The mechanism of oncogenesis by JNK will be tested by determining whether predicted DNA synthesis genes are expressed in tumor cells (Aim 3) and prostate tumor specimens (Aim 4) using established cDNA arrays. Microarrays will be constructed that utilize 35,000 NIH/CGAP sequence verified cancer expression library obtained by cloning from laser capture microdissection human tumor specimens. These studies test a novel hypothesis and test the utility of new inhibitors for the treatment of prostate carcinoma.

Jun 激酶/应激激活蛋白激酶通路 (JNK/SAPK) 被认为可以介导应激反应、存活、增殖和凋亡。 先前的研究（主要由 C. Der 及其同事以及本实验室参与的 M. Karin 进行）表明，激活的 ras 癌基因和相关癌基因对原代成纤维细胞的转化需要 JNK 途径。 我们已将 JNK 分析扩展到人类肿瘤细胞。 JNK 通常在多种人类肿瘤细胞中具有血清诱导性或组成型活性。特异性抑制可阻止肿瘤细胞生长。 此外，显性失活抑制剂 c-Jun (S63A、S73A) 的稳定表达可阻断 DNA 修复，并使人类肿瘤系（T98G 胶质母细胞瘤、A549 肺癌、MCF-7 乳腺癌和 PC3 前列腺癌）对顺铂的杀伤高度敏感。 PC3 细胞高达 9.7 倍。我们假设在癌症进展过程中，JNK 通路通常被选择为极大地增强 DNA 修复和合成，从而促进肿瘤发生。 为了测试这一点，我们开发并表征了与两个亚型家族 JNK1 和 JNK2 互补的高度特异性反义化合物。 对携带已建立的 PC3 异种移植物的无胸腺小鼠进行全身反义治疗可抑制生长 79%，优于顺铂 (49%)，并且联合反义治疗可促进高频率的完全消退。相反，PC3 细胞具有 100% 的致瘤性。 为了测试普遍性，发现另外 8 个前列腺细胞系的增殖与 JNK 活性的血清诱导水平成正比，范围超过 9 倍。 此外，反义JNK2而非反义JNK1几乎完全阻断所有细胞系的生长，与JNK活性成比例，强烈表明JNK2通常是血清刺激生长的前列腺癌细胞所必需的。 建议检验 JNK 在前列腺癌中具有致癌性的假设。 我们将在体外（目标 1）和体内（目标 2）测试 JNK 是否是前列腺癌细胞系增殖所必需的。 反义 JNK 的功效将在 TRAMP 模型中进行严格测试（目标 2）。 将通过使用已建立的 cDNA 阵列确定预测的 DNA 合成基因是否在肿瘤细胞（目标 3）和前列腺肿瘤样本（目标 4）中表达来测试 JNK 的肿瘤发生机制。 将利用从激光捕获显微切割人类肿瘤标本中克隆获得的 35,000 个 NIH/CGAP 序列验证的癌症表达文库构建微阵列。这些研究检验了一个新的假设并测试了新抑制剂治疗前列腺癌的效用。