1990 Fiscal Year Final Research Report Summary
Analysis of Differentially Expressed Gene in Myelogenous Leukemia
| Project/Area Number |
01570688
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Hematology
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| Research Institution | Kyushu University |
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Principal Investigator |
KUDO Jiro Kyushu University, Faculty of Medicine, Assistant, 医学部, 助手 (90148940)
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| Co-Investigator(Kenkyū-buntansha) |
ISHIBASHI Hiromi Kyushu University, Faculty of Medicine, Assistant, 医学部, 助手 (80127969)
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| Project Period (FY) |
1989 – 1990
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| Keywords | mitochondrial ND2 gene / leukemia / differential expression / thymosin beta_4 gene / differentiation |
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| Research Abstract |
We differentially screened 5,000 clones from a cDNA library of Acute Myelogenous Leukemia (AML) cell line HL60 using cDNA probes derived from normal granulocytes or from acute myelomonocytic leukemia cells, the objective being to identify genes preferentially expressed in myeloid lineage leukemic cells. One clone, corresponding to a mitochondrial DNA fragment, including NADH dehydrogenase submit 2 (ND2) gene, was selected for use as a probe. We examined expression of the ND2 gene in various leukemic cell populations and in normal peripheral blood cells. DNA-RNA hybridization studies revealed that ND2 messenger RNA (mRNA) was more highly expressed in AML cells than in other leukemic cells and normal peripheral granulocytes. The expression of ND2 mRNA decreased in HL60 cells several hours after treatment with phorbol myristate acetate (PMA), or with dimethylsulfoxide (DMSO). However, the ND2 gene expression did not depend on the growth-state of HL60 cells because the steady-state level of its expression was observed during transitions of growth. These results suggest that ND2 mRNA is involved in the maturation of myeloid cells and cellular differentiation in a lineage-preferential manner. A comparison of the nucleotide sequence of this clone with the documented human mitochondrial DNA sequence revealed several single-base substitutions, insertions and a 39-bases insertion. Moreover, we examined thymosin beta_4 (Tbeta_4) gene expression in leukemic cell lines. Different from ND2 gene, the Tbeta_4mRNA level decreased once during an early phose then increased to the high peak in HL60 cells treated with TPA or DMSO and in K562 cells treated with AraC or hemin.
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