2000 Fiscal Year Final Research Report Summary
Use of a Ligand Regulatable BCR/ABL Oncogene to Investigate the Pathogenesis of CML
| Project/Area Number |
11671011
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Hematology
|
|---|
| Research Institution | Kyoto Prefectural University of Medicine |
|---|
Principal Investigator |
OKUDA Keiko Kyoto Prefectural University of Medicine Medicine Instructor, 医学部, 助手 (70305572)
|
|---|
| Project Period (FY) |
1999 – 2000
|
| Keywords | Leukemia / Oncogene / Animal Model |
|---|
| Research Abstract |
The specific and final aims of this proposal are to understand how leukemic cells obtain a growth advantage over normal marrow cells, by generating a mouse model of ligand-regulated leukemia. For the purpose, a synthetic oncogene, EPOR/ABL, has been created by replacing the cytoplasmic domain of the erythropoietin receptor (EPOR) with c-ABL.My goal is to produce a mouse in which the ABL oncogene can be turned on and off, and then determine the in vivo consequences of this event. Practically, I focused my efforts on expressing the oncogene in primary marrow cells by infecting that with retroviruses carrying the chimeric oncogene. I obtained Packaging cell lines (Psy-cre, 293T and BOSC), then transfected with EPOR/ABL cDNA in vector pPL.While, there weren't any good subline to infect NIH3T3 nor Ba/F3 cells as a preliminary target. After swapping the cDNA to pLNCX vector, I could chose good subline at least to infect those cell lines, although, I haven't succeeded to express substantial amount of the chimeric protein in mouse bone marrow cells, using the same packaging cell line yet.
The optimal condition for marrow cells infection, is still under review.
In this term, I also studied on ABL related gene ARG.
The results I have observed on ARG so far, are as follows.
-The ARG tyrosine kinase is activated in the TEL/ARG oncogene.
-TEL/ARG induces anchorage-independent proliferation of Rat-1 cells.
-TEL/ARC induces factor-independent proliferation of Ba/F3 cells.
-STI571 inhibits tyrosine phosphorylation induced by TEL/ARG with an IC50 of 0.5μM
-STI571 inhibits proliferation and mobility of Ba/F3 cells transformed by TEL/ARG.
In addition to ABL, I'm going to use this ligand regulatable system to investigate the function of ARG tyrosine kinase and it's role in leukemogenesis.
|
Research Products
(4 results)
