Analysis of c-kit expression of human erythroleukemia cell line, HEL: clonal variation and relationship with erythroid and megakaryocytic phenotype.
The proto-oncogene c-kit encodes the receptor for a stem cell factor. We examined HEL, a human erythroleukemia cell line, in order to clarify the correlation between the c-kit receptor (KR) expression and lineage-specific phenotype. Although HEL cells are known to express KR, we found two relatively distinct HEL cell populations in terms of KR expression. We then subcloned HEL cell lines with clone sorting on the basis of KR expression and compared their various characteristics. The highly KR-expressing subline, HEL-P1, expressed a high level of glycophorin A (GPA), a known erythroid lineage marker. HEL-N1, in which most of the cells were KR-negative, showed a higher megakaryocytic lineage marker CD41b expression than HEL-P1. However, the expression of granulomonocytic lineage markers were not significantly different between the two subclones. Cell growth rate and cell cycle analysis also did not detect significant differences between the sublines. HEL-P1 cells gradually lost their KR expression in serum-containing culture, while the percentage of KR-positive HEL-N1 cells increased in serum-free culture. These observations indicate that KR expression was associated with the synchronous expression of GPA and inversely correlated with CD41b, and reversible transitions between KR-positive cells and KR-negative cells exist. We suggest that KR plays an important part in commitment of erythroid and megakaryocytic precursor cells.
Kubota
A
A
First Department of Internal Medicine, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
Okamura
S
S
Shimoda
K
K
Ikematsu
W
W
Otsuka
T
T
Niho
Y
Y
D015703:Antigens, CD / Q000378:metabolism*; D002453:Cell Cycle; D002469:Cell Separation; D002999:Clone Cells; D004912:Erythrocytes / Q000166:cytology; D005434:Flow Cytometry; D015870:Gene Expression; D006021:Glycophorins / Q000378:metabolism; D006801:Humans; D016130:Immunophenotyping; D004915:Leukemia, Erythroblastic, Acute / Q000235:genetics* / Q000473:pathology; D008533:Megakaryocytes / Q000166:cytology; D000090063:Proto-Oncogene Mas; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D019009:Proto-Oncogene Proteins c-kit; D012333:RNA, Messenger / Q000235:genetics; D012334:RNA, Neoplasm / Q000235:genetics; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics*; D016184:Receptors, Colony-Stimulating Factor / Q000235:genetics*; D014407:Tumor Cells, Cultured
D015703:Antigens, CD; D006021:Glycophorins; C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D016184:Receptors, Colony-Stimulating Factor; D019009:Proto-Oncogene Proteins c-kit; D020794:Receptor Protein-Tyrosine Kinases