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Journal of Histochemistry and Cytochemistry, Vol. 49, 565-572, May 2001, Copyright © 2001, The Histochemical Society, Inc.

ARTICLE

Oxygen Insensitivity of the Histochemical Assay of Glucose-6-phosphate Dehydrogenase Activity for the Detection of (Pre)Neoplasm in Rat Liver

Jonas S.S.G. De Jonga, Wilma M. Frederiksa, and Cornelis J.F. Van Noordena
a Academic Medical Center, University of Amsterdam, Department of Cell Biology and Histology, Amsterdam, The Netherlands

Correspondence to: Wilma M. Frederiks, Dept. of Cell Biology and Histology, Academic Medical Center, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands. E-mail: w.m.frederiks@amc.uva.nl

Oxygen insensitivity of the histochemical assay to detect glucose-6-phosphate dehydrogenase (G6PD) activity with NT as tetrazolium salt has been proved to be a powerful tool to discriminate various types of adenocarcinoma from normal tissues. Here we investigated whether this phenomenon can also be applied to differentiate between chemically induced hepatocellular (pre)neoplasms and normal liver tissue in rats. Residual activity (percentage of the amount of final reaction product that is generated in oxygen and that is generated in nitrogen) was 60% in (pre)neoplastic cells and 6% in normal liver parenchymal cells. This means that the oxygen insensitivity test is a useful tool to distinguish (pre)neoplasms from normal rat liver tissue. N-Ethylmaleimide, a blocker of SH groups, did not affect G6PD activity in (pre)neoplastic cells, whereas activity in normal cells was reduced by half. Therefore, the absence of essential SH groups in G6PD in (pre)neoplastic cells is held responsible for the oxygen insensitivity phenomenon. We conclude that oxygen insensitivity of the histochemical assay for G6PD activity is a fast, easy, and cheap tool to diagnose (pre)neoplasms in rat liver. Discrimination is likely to be based on altered properties of the enzyme in (pre)neoplastic cells. (J Histochem Cytochem 49:565–571, 2001)

Key Words: hepatocellular carcinoma, glucose-6-phosphate, dehydrogenase, enzyme histochemistry, neotetrazolium, diagnosis, oxygen insensitivity test, image analysis


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