Studies of myeloperoxidase gene expression at the cellular level by in situ hybridization.
Recent studies have demonstrated myeloperoxidase (MPO) gene expression during granulocytic differentiation. Since these studies have been done exclusively by Northern and dot blot analysis and frequently with mixed populations of cells, quantitative changes in gene expression for particular populations of cells are difficult to assess. We therefore examined MPO expression at the cellular level in various normal and malignant hematopoietic cells by the in situ hybridization (ISH) technique. Using this approach, we demonstrated that inducing the promyelocytic HL-60 cell line to differentiate along either monocytic or granulocytic pathways decreases MPO mRNA expression. Similarly, when ISH was performed on normal bone marrow, relatively high levels of MPO mRNA were detected in myeloblasts, promyelocytes, and early eosinophilic precursors, whereas the expression was markedly decreased in more advanced stages of myeloid differentiation. These findings agree with the known decrease in MPO protein synthesis observed during granulocytic differentiation and suggest that regulation of MPO protein synthesis occurs at the level of MPO mRNA expression. We conclude by showing that ISH can detect MPO mRNA in myeloblasts of patients with acute leukemia and can be a potentially useful technique in the study of myeloid differentiation in acute leukemias.
Zaki
S R
SR
Division of Immunologic, Oncologic, and Hematologic Diseases, Centers for Disease Control, Atlanta, Georgia 30333.
Austin
G E
GE
Swan
D C
DC
Hooper
W C
WC
Greer
P W
PW
Evatt
B L
BL
Chan
W C
WC
D000293:Adolescent; D000328:Adult; D000368:Aged; D001345:Autoradiography; D001853:Bone Marrow / Q000201:enzymology; D002454:Cell Differentiation; D002460:Cell Line; D005260:Female; D015870:Gene Expression; D006410:Hematopoiesis; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000201:enzymology / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D009693:Nucleic Acid Hybridization; D009195:Peroxidase / Q000096:biosynthesis / Q000235:genetics*; D010641:Phenotype; D012333:RNA, Messenger / Q000378:metabolism*
D012333:RNA, Messenger; D009195:Peroxidase