Antisense down-regulation of 5-FU related enhances cytotoxicity of 5-FU in human lung cancer
| Project/Area Number |
13671237
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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 |
General surgery
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| Research Institution | THE University of Tokushima |
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Principal Investigator |
KONDO Kazuya University of Tokushima, school of medicine, Assistant Professor, 医学部, 講師 (10263815)
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| Co-Investigator(Kenkyū-buntansha) |
MIYOSHI Takanori University of Tokushima, university hospital, Assistant, 医学部附属病院, 助手 (20346612)
MONDEN Yasumasa University of Tokushima, school of medicine, Professor, 医学部, 教授 (60028628)
滝沢 宏光 徳島大学, 医学部・附属病院, 医員
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2002: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2001: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | antisense / oligodeoxynucleotide / thymidylate synthase / liposome / lung cancer / 5-FU / thymidylate synthase / antisense oligonucleotide / HVJ-リポソーム |
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| Research Abstract |
Thymidylate synthase (TS), the key enzyme in de novo synthesis of thymidine, is an important target for antitumor chemothrapy of 5-FU. We used antisense oligodeoxynucleotide (ODN) to down-regulate the expression of TS. We made 5 antisense ODNs which is complementary to TS Mrna, and one random ODN. Transfection of ODNs was performed using HVJ-liposome. After incubating human lung cancer cell lines (Ma10 and Ma25) and antisense ODNs with HVJ-liposome, the TS expression of cell lines was examined by RT-PCR method and TS activity was measured. TS expression was repressed from 50% to 20% by incubaating cell lines and antisense ODNs and TS expression. It was impossible that antisense ODNs of TS inhibited the expression of TS. Chu e al. reported that the TS protein itself binds to the T Mrna both at the translational start site and in the coding region, inhibiting translational processing of the message.
Next year, we examined whether CDDP (anticancer drug) conjugated by liposome (liposome-CDDP) enhance anticancer effect. The IC50 of CDDP was 40ug/ML in human lung cancer cell line Ma44-3 in vitro. Meanwhile, the IC50 of liposome-CDDP was 10ug/ML for 48 hr incubation. And it was 1.5ug/ML for 96 hr incubation. When thecubation time becomes longer, the IC50 of liposome-CDDP was lower. The use of Rhodamine-labeled liposome-CDDP revealed that the lipsome-CDDP attached the cytoplasm of cancer cells in vitro. Rhodamine-labeled liposome-CDDP revealed that the liposome-CDDP attached the cytoplasm of cancer cells in vitro. Rhodamine-labeled liposome-CDDP was administrared in SCID mice using a nebulizer (through airway). Rhodamine-labeled liposome was observed in the lung of the mice. These results demonstrate that the administration of liposome-CDDP through airway may become a therapy for lung cancer.
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Report
(3 results)
Research Products
(13 results)
