J M Cline1, D E Thrall, G L Rosner, J A Raleigh. 1. Department of Comparative Medicine, Bowman Gray School of Medicine of Wake Forest University, Winston-Salem, NC 27157-1040.
Abstract
PURPOSE: The purpose of this study was to identify the prevalence and distribution of hypoxic tumor cells in spontaneous canine tumors, and to relate these parameters to various tumor and patient characteristics, such as tumor volume, tumor type, or tumor location. METHODS AND MATERIALS: Hypoxic tumor cells were labeled in vivo in 32 primary malignant canine tumors by bioreductive binding of the nitroimidazole hypoxia marker CCI-103F. CCI-103F was given at 40 mg/kg i.v. Tumors were completely excised, and CCI-103F adducts were detected in histologic sections (mean, 138 sections-per-tumor) by peroxidase-antiperoxidase immunostaining. Area fraction (area labeled/total area examined) of labeled regions was measured via computer assisted image analysis. In tumors with a volume < 100 cm3, each cubic centimeter of tumor was examined; in larger tumors 100 randomly selected 1 cm3 samples were examined. RESULTS: There were 13 soft-tissue sarcomas, 11 mast-cell tumors, five carcinomas, two lymphosarcomas, and one melanoma. Tumors varied from < .001 to > 2000 cm3. Labeled cells were present in 31 of 32 canine tumors examined, and varied between 0 and 35%. Mean (+/- SD) % label was 12.2% +/- 16.7%; 13 of the 32 dogs had % labeled area < 5.0%. The area fraction was not related to tumor site, tumor type, tumor volume, presence and degree of necrosis or tumor grade. Dog characteristics such as sex, age, and body size did not affect the degree of labeling of tumors. CCI-103F adducts were randomly distributed grossly, and at the microscopic level were not found near blood vessels or regions containing mitoses. Labeling was seen in a variety of normal tissues; not all binding in normal tissues could be attributed to hypoxia. CONCLUSION: CCI-103F labeling of hypoxic regions in tumors provides a nonradioactive method of detecting nitroimidazole adducts at the cellular level, and allows concurrent histologic examination. The pattern of labeling is consistent with detection of hypoxic tumor cells arising from oxygen diffusion limitations. This method may have clinical applicability in the detection of tumor hypoxia.
PURPOSE: The purpose of this study was to identify the prevalence and distribution of hypoxic tumor cells in spontaneous caninetumors, and to relate these parameters to various tumor and patient characteristics, such as tumor volume, tumor type, or tumor location. METHODS AND MATERIALS: Hypoxic tumor cells were labeled in vivo in 32 primary malignant caninetumors by bioreductive binding of the nitroimidazolehypoxia marker CCI-103F. CCI-103F was given at 40 mg/kg i.v. Tumors were completely excised, and CCI-103F adducts were detected in histologic sections (mean, 138 sections-per-tumor) by peroxidase-antiperoxidase immunostaining. Area fraction (area labeled/total area examined) of labeled regions was measured via computer assisted image analysis. In tumors with a volume < 100 cm3, each cubic centimeter of tumor was examined; in larger tumors 100 randomly selected 1 cm3 samples were examined. RESULTS: There were 13 soft-tissue sarcomas, 11 mast-cell tumors, five carcinomas, two lymphosarcomas, and one melanoma. Tumors varied from < .001 to > 2000 cm3. Labeled cells were present in 31 of 32 caninetumors examined, and varied between 0 and 35%. Mean (+/- SD) % label was 12.2% +/- 16.7%; 13 of the 32 dogs had % labeled area < 5.0%. The area fraction was not related to tumor site, tumor type, tumor volume, presence and degree of necrosis or tumor grade. Dog characteristics such as sex, age, and body size did not affect the degree of labeling of tumors. CCI-103F adducts were randomly distributed grossly, and at the microscopic level were not found near blood vessels or regions containing mitoses. Labeling was seen in a variety of normal tissues; not all binding in normal tissues could be attributed to hypoxia. CONCLUSION:CCI-103F labeling of hypoxic regions in tumors provides a nonradioactive method of detecting nitroimidazole adducts at the cellular level, and allows concurrent histologic examination. The pattern of labeling is consistent with detection of hypoxic tumor cells arising from oxygen diffusion limitations. This method may have clinical applicability in the detection of tumor hypoxia.