| Project/Area Number |
05670493
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
Gastroenterology
|
|---|
| Research Institution | Nara Medical University |
|---|
Principal Investigator |
KURIYAMA Shigeki Nara Medical University, Medicine, Research Associate, 医学部, 助手 (50244710)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
YOSHIKAWA Masahide Nara Medical University, Medicine, Research Associate, 医学部, 助手 (50230701)
TSUJII Tadasu Nara Medical University, Medicine, Professor, 医学部, 教授 (30075064)
|
|---|
| Project Period (FY) |
1993 – 1994
|
| Project Status |
Completed (Fiscal Year 1994)
|
| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1994: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1993: ¥1,400,000 (Direct Cost: ¥1,400,000)
|
|---|
| Keywords | Hepatocellular carcinoma / Gene therapy / Retroviral vector / Gene transfer / Albumin gene / Promoter / beta-Galactosidase / レトロウイルスベクター / 組織特異性 / 肝細胞 |
|---|
| Research Abstract |
For in vivo gene therapy against cancer, it is indispensable to direct the expression of exogenous genes exclusively to cancer cells. We have constructed a recombinant retroviral vector in which a tissue-specific regulatory element is used as an internal promoter. It contains the murine albumin enhancer and promoter as an internal promoter and the lacZ gene coding bacterial beta-galactosidase as a reporter between the two Moloney murine leukemia virus long terminal repeats. This vector was introduced into ecotropic retroviral phi2 packaging cells to produce recombinant retroviral particles. Various cell lines were infected with the recombinant retrovirus to assess its tissue specificity in vitro. Expression of the lacZ gene was detected solely in hepatoma cell lines but not in fibroblasts nor in B lymphoma cells. In vivo infection was carried out to evaluate tissue specificity of this system. The recombinant retrovirus was injected into a murine subcutaneous hepatoma and expression of the lacZ gene was observed in hepatoma cells but not in surrounding connective tissues. Then the retrovirus was injected into murine livers and no expression was detected in normal hepatocytes. The retrovirus was again injected in to livers of the partially hepatectomized mice, resulting in the gene expression in only a few regenerating hepatocytes. Ratio of expression of the exogenous gene was, however, much lower than that in subcutaneous hepatoma. Finally, the gene expression of higher efficiency was confirmed as the retroviral infections were repeated. These results indicate that repeated gene transfer by means of the recombinant retrovirus which contains a tissue-specific regulatory element as an internal promoter should possess high potential for selective elimination of hepatoma cells in vivo.
|
