| Project/Area Number |
10470251
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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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 | Tohoku University |
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Principal Investigator |
SUNAMURA Makoto (1999-2001) Tohoku Univ. Hospital, Lecturer, 医学部・附属病院, 講師 (10201584)
小針 雅男 (1998) 東北大学, 医学部, 助教授 (30170369)
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| Co-Investigator(Kenkyū-buntansha) |
FURUKAWA Toru Tohoku Uni Grad Sch Med, Research Associate, 大学院・医学系研究科, 助手 (30282122)
TAKEDA Kazunori Tohoku Uni Grad Sch Med, Ass Prof, 大学院・医学系研究科, 助教授 (20171639)
MATSUNO Seiki Tohoku Uni Grad Sch Med, Professor, 大学院・医学系研究科, 教授 (80004737)
SIBUYA Kazuhiko Tohoku Uni Hospital, Research Associate, 医学部・附属病院, 助手 (70260429)
島村 弘宗 東北大学, 医学部・附属病院, 助手 (70312585)
砂村 眞琴 東北大学, 医学部・附属病院, 助手 (10201584)
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| Project Period (FY) |
1998 – 2000
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥12,300,000 (Direct Cost: ¥12,300,000)
Fiscal Year 2000: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1999: ¥5,200,000 (Direct Cost: ¥5,200,000)
Fiscal Year 1998: ¥4,100,000 (Direct Cost: ¥4,100,000)
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| Keywords | pancreatic cancer / gene therapy / uracil phosphoribosyl transferase / 5FU / angiogenesis / soluble VEGF receptor / flt 1 / ハイブリッド型リポソームベクター / 癌 / P53遺伝子 / アデイウイルス / UPRT / UPRT,CD, / 増殖型アテツウイルス / 変異型アデノウィルスベクター / UPRT遺伝子 |
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| Research Abstract |
E. coli uracil phosphoribosyltransferase (UPRT) catalyzes the synthesis of FUMP from 5-FU, resulting in increased production of FdUMP, which induces cytotoxic effect by inhibiting the function of thymidylate synthesis. We constructeded UPRT expressing mutant adenovirus vector (AxElAdB-UPRT) that replicates preferentially in tumor cells without p53 function. Pancreatic cancer cells lacking p53 function were transfected with either UPRT or lacZ gene using adenovirus vectors (Ax-UPRT and Ax-lacZ) and cultured in medium containing various concentration of 5-FU. Therapeutic advantage of AxE1AdB-UPRT/5-FU treatment was evaluated using intraperitoneal disseminated ASPC-1 tumor mice model. The mice were treated with intraperitoneal injection of vectors 10 days after tumor inoculation, followed by the administration of 5-FU(10 mg/Kg) for 7days. Mice were sacrificed and tumor weight was measured 4 weeks later. The sensitivity against 5-FU of 5 pancreatic cancer cells increased significantly (7.1
… More
-72 fold) after the transfection of UPRT gene. Tumor weight in Ax-UPRT/5-FU group (0.24 g) was significantly reduced as compared to Ax-lacZ/5-FU group (0.56 g) (P<0.001). Furthermore, that of AxElAdB-UPRT/5-FU group was significantly decreased (0.15 g).(P=0.042 vs Ax-UPRT/5-FU group). Furthermore, the adverse effect of FdUMP on alimentary tract was diminished because of the decreasd dosage of 5-FU.
The growth of a tumor and its ability to metastasize is angiogenesis-dependent. Angiogenesis is one of the crucial steps in tumor's transition from a small, harmless cluster of mutated cells to a large, malignant growth, capable of spreading to other organs throughout the body soluble. We transfected Panc-1 cells with soluble VEGF receptor (flt1) and as control with the lacZ gene using the adenovirus vectors Adsflt and AdlacZ, respectively. Adsflt or AdlacZ (3.5X10^7 PFU) or 100 μl of PBS were injected directly into tumors daily from day 8 through day 12 after the implantation. Furthermore, we implanted in dorsal skinfold chamber the wild-type Panc-1 and its transfectants and monitored the tumor angipgenesis using in vivo microscope system. VEGF levels in conditioned media were higher in all cell lines (mean, 1969 pg/ml ; Panc-1, 980 pg/ml), compared to the level in media of HUVEC (8 pg/ml). Expression of VEGF mRNA was detected in the all cell lines by RT-PCR and Northern blot analysis and it was confirmed by DNA sequencing. Tumor growth rate of Adsflt-infected cells was lower than those of AdlacZ group and no treatment group, and the difference was statistically significant. Wild type Panc-1 and the LacZ transfectant implantation prompted strong tumor angiogenesis as observed in skinfold chamber, whereas soluble flt-1 transfectants failed to exert such an effect. Less
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