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...

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Bibliographic Details
Published in:The journal of histochemistry and cytochemistry 2007-02, Vol.55 (2), p.175-181
Main Authors: Frederiks, Wilma M, Vreeling-Sindelarova, Heleen, Van Noorden, Cornelis J.F
Format: Article
Language:English
Subjects:
Animals
Catalase - metabolism
Cell Line, Tumor
Colonic Neoplasms
Ethylmaleimide - pharmacology
Glucosephosphate Dehydrogenase - metabolism
Hepatocytes - enzymology
Histocytochemistry
Humans
In Vitro Techniques
Liver Neoplasms
Male
Oxygen - metabolism
Peroxisomes - enzymology
Peroxisomes - ultrastructure
Rats
Rats, Wistar
Sulfhydryl Reagents - pharmacology
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Summary: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.
ISSN:0022-1554
1551-5044
DOI:10.1369/jhc.6A7068.2006