1995 Fiscal Year Final Research Report Summary
Proliferation of leukemic cells by retrovirus mediated gene transfer
| Project/Area Number |
05670912
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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 |
OTSUKA Teruhisa Kyushu University, Medicine, Assistant professor, 医学部, 講師 (20185317)
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| Co-Investigator(Kenkyū-buntansha) |
IWASAKI Hiromi Kyushu University, Medicine, Medical Staff, 医学部, 医員
OHNO Yuju Kyushu University, Medicine, Medical Staff, 医学部, 医員
SHIGEMATSU Hirokazu Kyushu University, Medicine, Medical Staff, 医学部, 医員
ARIMA Fumitou Kyushu university, Medicine, Medical Staff, 医学部, 医員
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| Project Period (FY) |
1993 – 1995
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| Keywords | retrovirus / leukemia / autocrine proliferation |
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| Research Abstract |
The mechanisms of proliferation of leukemic cells were investigated by retrovirus-mediated gene transfer. Initially, we have constructed theretroviral vector which was inserted the cDNA for hematopoietic growth factors. We tried to construct the G-CSF, GM-CSF, IL-3 and IL-6 secreting retroviral vectors. These vectors were transfected to the ecotropic packaging cell line.phi2 and were selected with G418. Then, the supernatant of phi2 cells were infected to the amphotropic ppackging cell line, PA317&Am12. We have successfully constructed the G-CSF, GM-CSF,IL-3 and IL-6 secreting retroviral vectors with hight viral and growth factor titer. Using these retroviral vectors, we initially infected the factor dependent murine leukemic cell lines. When we, infected the IL-6 dependent cell line, B9・8, wiht IL-6 producing retrovirus, this B9・8 cells proliferated autonomously. However, when we infected the control retrovirus, these cells showed the IL-6 dependent and they died by apoptosis without IL-6. We confirmed the gene integration by PCR , method. Next, we infected the G-CSF dependent cell line NFS-60 with G-CSF producing retrovirus. These infected cells also showed autonomous growth. Interestingly, these cells died by apoptosis when these infected cells reached to the confluent in culture. Finally, we tried to infect the hematopoietic growth factor producing retrovirus to the de novo myelogenous leukemic cells and myeloma cells to get the stable cell line. So far we have not succeeded the get the stable cell line after infection.
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