Cytogenetic analysis of malignant cells from patients with leukemia have recently been shown to be of significant value in determining prognosis, predicting response to therapy, and in developing a better insight into the biology of the disease. Similar correlations in human solid tumors have been hampered by difficulties in obtaining karyotypes directly from tumor specimens. Much of the cytogenetic data obtained so far have been obtained from derived cell lines and from metastatic effusions where selective cell growth, culturing artifacts, or clonal evolution cloud the original cytogenetic alterations. The purpose of this proposal is to perform direct cytogenetic analysis on fresh and cultured tumor specimens and normal tissue specimens using the technique of premature chromosome condensation. With this technique, the tumor or normal cell populations are fused with mitotic cell populations, resulting in the formation of prematurely condensed chromosomes (PCC). These preparations can then be banded and the karyotypes established. Thus the tumor or normal cell population need not be cycling nor is there a requirement for their growth in vitro. The objectives of this proposal are to determine whether specific cytogenetic changes are associated with lung cancer and head and neck cancer, and determine whether these changes are of prognostic value. In addition, since both diseases are associated with a carcinogen-induced """"""""field of cancerization, normal adjacent tissue wil be analyzed to determine whether cytogenetic changes are associated with the risk of recurrence. The direct karyotypes found by the PCC technique will be compared with that obtained by conventional mitotic cytogenetic analysis to determie whether clonal selection occurs with in vitro culture. The PCC technique will also be improved (e.g. selection for tumor cell fusion and the use of second tumor-specific markers to identify the fused cells of interest) to enhance the ability to characterize the cells of interest from a heterogeneous population. It is hoped that this study will provide a technical prototype for the cytogenetic analysis of all solid tumors, detect karyotypic changes associated with the multistep malignant process, and identify karyotypic changes associated with resistance to therapy within the human tumor setting.
