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American Journal of Pathology, Vol 142, 307-317, Copyright © 1993 by American Society for Investigative Pathology

REGULAR ARTICLES

Interphase cytogenetics in paraffin sections of lung tumors by non- isotopic in situ hybridization. Mapping genotype/phenotype heterogeneity

SY Kim, JS Lee, JY Ro, ML Gay, WK Hong and WN Hittelman
Department of Medical Oncology, University of Texas M. D. Anderson Cancer Center, Houston 77030.

To better understand the relationship between specific chromosome changes found in human lung tumors and their phenotypic consequences at the tissue level, an in situ hybridization procedure was optimized for use on formalin-fixed paraffin-embedded tissue sections of human lung tumors. Pretreatment heating of sections, pepsin concentration, duration of pepsin treatment, hybridization conditions, and posthybridization washing conditions were varied to determine optimum conditions. The deparaffinized sections were stained with centromeric probes for chromosomes 7 and 17, and a chromosome index for each tumor was derived by dividing the mean number of chromosome signals found on the tumor cells by the mean number of chromosome signals on normal cells (lymphocytes and fibroblasts) in the same section. This chromosome index was then compared with the DNA index determined in an adjacent section by Feulgen staining followed by image analysis quantitation. The chromosome index correlated well with the DNA index, but in some cases, chromosome 7, or 17 in other cases, was either over- or under-represented compared with the corresponding DNA index. In addition, chromosome and DNA alterations were shown to be differentially expressed within the same tissue section, correlating with a change in tumor differentiation status. These results suggest that in situ hybridization will prove to be an important tool for determining the underlying genetic basis for tissue phenotype heterogeneity by allowing genetic determinations to be made on paraffin- embedded tissue sections where tumor histological architecture is preserved.

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