| Project/Area Number |
09670557
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Gastroenterology
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| Research Institution | Nagasaki University |
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Principal Investigator |
NAKAO Kazuhiko Nagasaki University Health Research Center, Lecturer, 保健管理センター, 講師 (00264218)
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| Co-Investigator(Kenkyū-buntansha) |
NAKATA Keisuke Nagasaki University Department of Medicine, Assistant professor, 医学部, 助教授 (40217740)
馬詰 裕之 長崎大学, 医学部附属病院, 医員
有馬 哲彦 長崎大学, 医学部附属病院, 医員
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 1998: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1997: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Gene therapy / Hepatocellular carcinoma / alpha-fetoprotein (AFP) / Recombinant retrovirus / 組換えレトロウィルス / 肝癌 / レトロウイルスベクター / ヒトAFP |
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| Research Abstract |
The alpha-fetoprotein (AFP) gene is reactivated in hepatocellular carcinoma.. We have previously reported that a retrovirus vector (LNAFO.3TK) carrying a herpes simplex virus thymidine kinase gene regulated by the 0.3-kb human AFP promoter provides ganciclovir (GCV)-mediated cytotoxicity in AFP-producing hepatoma cells parallel with the ability of AFP production.
In the present study, the in vivo effect of retrovirus-mediated gene therapy for human hepatoma cells was examined. High AFP-producing human hepatoma cells (HuH-7) was infected with LNAFO.3TK and implanted into subcutaneous of athymic mice. GCV treatment resulted in pronounced growth inhibition of the virus-infected HuH-7 xenograft in mice, but did not affect growth of parental xenograft.
To improve the efficacy of retrovirus-mediated gene therapy for the intermediate and low AFP-producing hepatoma cells, new recombinant retrovirus vector (LNAFO.3E+TK) was constructed, in which human AFP enhancer region was directly linked to 0.3-kb AFP promoter of LNAEO.3TK.In the intermediate and low AFP-producing human hepatoma cells PLC/PRF/5 and huH/cl.2, respectively, LNAFO.3E+TK sensitized these cells to GCV in vitro. In vivo model using athymic mice harboring PLC/PRF/5 cells, GCV treatment resulted in more pronounced growth inhibition in the LNAFO.3E+TK virus-infected cells than in LNAFO.3TK virus-infected cells.
It is reported that a G o A substitution in the human APP promoter is associated with hereditary persistence of APP, therefor this substitution was generated in LNAFO.3TK to construct LNAFO.3MTK.LNAFO.3MTK infection into PLQTPRF/5 and huH/cl.2 cells also showed more pronounced sensitivity to GCV treatment than LNAFO.3TK infection.
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