MOLECULAR CHANGES DURING PROSTATE CARCINOMA PROGRESSION

前列腺癌进展过程中的分子变化

• 批准号: 6300434
• 负责人: RAYMOND B NAGLE
• 金额: $ 13.98万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6300434
• 关键词: SCID mouse; antidromic impulse; biological signal transduction; carcinogenesis; cell adhesion; cell growth regulation; cell line; cell migration; complementary DNA; gene expression; genetic translation; human tissue; immunoprecipitation; in situ hybridization; integrins; laminin; membrane proteins; messenger RNA; molecular oncology; neoplasm /cancer invasiveness; neoplastic process; nucleic acid sequence; polymerase chain reaction; posttranscriptional RNA processing; prostate neoplasms; protein biosynthesis; protein degradation; protein purification; scanning electron microscopy; tissue /cell culture; transmission electron microscopy; western blottings

The transition from prostatic intrepithelial neoplasia (PIN) to invasive carcinoma is characterized by a loss of two major regulatory proteins, beta4 integrin and its ligand, laminin 5. We hypothesize that the mechanism for the loss of laminin 5 is the presence of a post-transcriptional defect. The failure of laminin 5 assembly causes instability of beta4 which is lost due to degradation. The loss of these regulatory proteins destabilizes the normal signaling pathway of the cells contributing to prostate tumor progression. We will examine this hypothesis by using the LNCaP cell line which, like the primary prostate carcinomas, expresses the three laminin subchain mRNAs but fails to assemble the fully functioning heterotrimeric laminin 5 protein. In Aim 1 we will examine the three laminin 5 subchain mRNAs for mutations by sequencing fragments of the cDNA products produced by RT-PCR. The mRNAs will also be tested for their in vitro translational capacity. In Aim 2 we will test the effect the purified laminin 5 on the synthesis, degradation, and cytoplasmic membrane stability of beta4 integrin. We will also study the ability of purified laminin 5 to cause signal transduction. These experiments will utilize pulse labeling or surface biotinylation followed by immunoprecipitation, and in the cause of signal transduction, Western blotting with anti-phosphotyrosine, Sh2, and GrB2 specific antibodies. The functional effect of laminin 5 on prostrate cell growth, adhesion, migration, and tumorigenicity will also be studied in Aim 3. These studies will be carried out using quantitative assays of adhesion and migration established in our laboratory. In Aim 4 we will test the clinical relevance of the persistence of alpha 6 and alpha 3 integrins in the absence of beta4.

前列腺上皮内瘤变（PIN）向侵袭性瘤变的转变