To further improve the survival of patients with non-small cell lung cancer (NSCLC), achievement of a thorough understanding of its biology must occur. In addition, clinicians willing and able to translate basic science advances to the patient must exist. This proposal has as main objective to provide skills and knowledge in basic sciences to a clinical investigator; key ingredients for a successful translational research career. The frequency of abnormalities in the expression of the pl6 suppressor gene in NSCLC, make the study of this gene a natural target to accomplish our objective. We propose the development of a transgenic mouse model of NSCLC to study in vivo the importance of the expression of pl6 in this neoplasia. Since an example of inactivation of this gene is hypermethylation, a mechanism susceptible to pharmacological manipulations, we will study the importance of this mechanism in the inactivation of pl6 in patients with NSCLC. In order to develop a NSCLC mouse model, a chimeric gene comprising an activated K-ras under the control of the human surfactant protein C (SP-C) transcriptional region will be constructed, and transgenic mice harboring this gene will be developed. Studies of apoptosis, flow cytometry, and expression of cyclin D1, cyclin dependent kinases, Rb, p16 and p53 will be studied. Subsequently, the SP-C/K-ras transgenic mice and pl6 knockout mice will be interbred. The effects of the absence of pl6 on histology, incidence, median age of tumor onset will be examined, as well as cell cycle and rate of apoptosis in those tumors. In order to evaluate the importance of methylation as a mechanism of p16 inactivation in NSCLC patients, tumor samples of 50 NSCLC patients undergoing resections or biopsies, will be obtained. Cell blocks of 50 other NSCLC patients will be also analyzed. Methylation-specific PCR (MSP) will be used to determine methylation in fresh tumor samples and cell blocks. Levels of cytosine-methyl transferase in peripheral blood cells, and levels of Fhgb will also be obtained in the patients undergoing resections or biopsies. A period of didactic and practical learning in molecular biology and its techniques, as well as in the basics of transgenic mice and husbandry, followed by the research plan detailed above, shall prepare the clinical investigator to effectively target clinical trials to translational work, and shall provide him with good insight into the biology of NSCLC.
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| Duan, Wenrui; Ding, Haiming; Subler, Mark A et al. (2002) Lung-specific expression of human mutant p53-273H is associated with a high frequency of lung adenocarcinoma in transgenic mice. Oncogene 21:7831-8 |
