One long-term objective of this project is to provide information on chromosome abnormalities in gynecologic tumors, and to find """"""""specific"""""""" and """"""""common"""""""" abnormalities in these tumors. Specific chromosome abnormalities now have diagnostic implications and the common ones may provide significant independent prognostic information. Also, site-specific chromosomal changes will identify the chromosomal segments containing genes which may be crucial to the malignant development. The intention is to determine the usefulness of chromosome studies in gynecologic tumors when sufficient data can be accumulated. Surgically removed tumor materials from reproductive organs, as well as effusions, are available for this study. Cytogenetic findings will add to and compare with the existing computerized information on the patients' clinical status and the course of their diseases. Study of both tumor tissue and effusion offers the opportunity of comparing the karyotypic changes in primary and metastatic sites. Caner of the uterine cervix is preceded by a spectrum of precancerous changes. Cytogenetic findings of various lesions of the cervix could be most valuable in the understanding of chromosomal changes in tumorigenesis. Each specimen is routinely processed for chromosome analysis, both directly and after short-term culture. Defined medium containing a fibroblast inhibitor, e.g. spermine, is used for the development of cell lines in culture. Exogeneous and autologous feeder layers will be used to initiate tumor cell growth when they are needed. Chromosome preparation is carried out according to standard protocols. Cell synchronization procedures are sometimes performed to enhance the yield of mitosis. When a specific chromosome rearrangement is found to be associated with a particular tumor, we will employ in situ chromosomal hybridization. Southern analysis, and pulsed field gradient electrophoresis to study the involvement of candidate gene(s) in this disorder. Amplification of cellular oncogenes could be a prognostic index in cancers. We plan to identify the gene(s) whose amplification is closely related to certain gynecologic tumors. Extensive in situ hybridization analyses of specially fixed and processed tumor tissues with numbers of growth related gene probes are performed and will help to select the most appropriate gene(s) for DNA analysis. Conclusive results of DNA analysis will be followed by the characterization of possible abnormal transcripts by means of Northern and nuclease S1 protection analysis.
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