| Project/Area Number |
17591409
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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 |
Digestive surgery
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| Research Institution | Ehime University |
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Principal Investigator |
HONDA Kazuo Ehime University, Graduate School of Medicine, Associate Professor, 大学院医学系研究科, 助教授 (00209321)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2006: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2005: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Chorangiocarcinoma / suicide gene / adenovector / UPRT / HSV-tk / tumor-specific replication |
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| Research Abstract |
The prognosis of bile duct cancer is considerably poor because the resection rate is low and cholangiocarcinoma is rather tolerant to chemotherapy and radiotherapy. We produced a recombinant adenovector (AxCAUT) with the uracil phosphoribosiltransferase (UPRT) gene of eschelichia coli and the thymidine kinase gene of Herpes simplex virus (HSV-tk), and the anti-tumor effect with 5-fluorouracil (5-FU) and ganciclovir (GVC) on human cholangiocarcinoma cells with a mutation of p53 (HuCCT1) was evaluated in vitro and in vivo. Furthermore, a conditionally replicating adenovirus with the deletion of E1B-55kD (AxE1CAUT) recombined with both the UPRT gene and the HSV-tk gene was produced and the anti-tumor effect was estimated. The drug sensitivity of HuCCT1 cells to 5-FU or GVC became higher with the increase of multiplicity of infection (M0I) of AxCAUT. The anti-tumor effect was evident when 5-FU and GVC were given at the same time. The sensitivity of AxE1CAUT-infected HuCCT1 cells to 5-FU/GV
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C increased in proportion to MOI. Subcutaneous tumors of nude mice infected with AxCAUT by direct injection showed higher response to 5-FU/GVC than 5-FU or GVC alone, but there was no significant difference between AxCAUT and AxE1CAUT. However, AxE1CAUT with 5-FU/GVC showed a significantly better survival rate than AxCAUT in a peritoneal dissemination model infected by intra-peritoneal injection of the adenovectors. In conclusion, the double suicide gene therapy was effective on cholangiocarcinoma cells. When direct administration of adenovectors into tumor is impracticable, oncolytic gene therapy combined with double suicide genes is a promising method, for example, in case of administration of the vector into the bile in the bile duct for the treatment of bile duct cancer.
The effect of the gene transfer of UPRT with AxCAUT to cholangiocarcinoma cells on the expression of thymidilate synthase (TS) was evaluated using real-time PCR. The level of TS was unaffected by the overexpression of UPRT. Less
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