Loss of peroxisomes causes oxygen insensitivity of the histochemical assay of glucose-6-phosphate dehydrogenase activity to detect cancer cells.
Oxygen insensitivity of carcinoma cells and oxygen sensitivity of non-cancer cells in the histochemical assay of glucose-6-phosphate dehydrogenase (G6PD) enables detection of carcinoma cells in unfixed cell smears or cryostat sections of biopsies. The metabolic background of oxygen insensitivity is still not understood completely. In the present study, rat hepatocytes, rat hepatoma cells (FTO-2B), and human colon carcinoma cells (HT29) were used to elucidate these backgrounds. The residual activity in oxygen was 0%, 55%, and 80% in hepatocytes, hepatoma cells, and colon carcinoma cells, respectively. N-ethylmaleimide (NEM), a blocker of SH-groups, did not affect G6PD activity in both carcinoma cell types but reduced G6PD activity in hepatocytes by 40%. Ultrastructural localization of G6PD activity was exclusively in the cytoplasm of carcinoma cells, but in hepatocytes both in cytoplasm and peroxisomes. NEM abolished peroxisomal G6PD activity only. Histochemical assay of catalase activity demonstrated absence of peroxisomes in both carcinoma cell lines. It is concluded that absence of SH-sensitive G6PD activity in peroxisomes in cancer cells is responsible for the oxygen-insensitivity phenomenon.
Frederiks
Wilma M
WM
Department of Cell Biology and Histology, Academic Medical Center, Meibergdreef 15 1105 AZ, Amsterdam, The Netherlands. w.m.frederiks@amc.uva.nl
Vreeling-Sindelárová
Heleen
H
Van Noorden
Cornelis J F
CJ
D000818:Animals; D002374:Catalase / Q000378:metabolism; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms; D005033:Ethylmaleimide / Q000494:pharmacology; D005954:Glucosephosphate Dehydrogenase / Q000378:metabolism*; D022781:Hepatocytes / Q000201:enzymology; D006651:Histocytochemistry; D006801:Humans; D066298:In Vitro Techniques; D008113:Liver Neoplasms; D008297:Male; D010100:Oxygen / Q000378:metabolism*; D020675:Peroxisomes / Q000201:enzymology* / Q000648:ultrastructure; D051381:Rats; D017208:Rats, Wistar; D013439:Sulfhydryl Reagents / Q000494:pharmacology
D013439:Sulfhydryl Reagents; D005954:Glucosephosphate Dehydrogenase; D002374:Catalase; D005033:Ethylmaleimide; D010100:Oxygen