Mechanism of the regulation of thymidylate synthase levels associated with response to 5-fluorouracil in human gastric cancer cells
| Project/Area Number |
20591562
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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 | Chiba University |
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Principal Investigator |
NABEYA Yoshihiro Chiba University, 医療局・消化器外科, 主任医長 (40322028)
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| Co-Investigator(Kenkyū-buntansha) |
HIWASA Takaki 千葉大学, 大学院・医学研究院遺伝子生化学, 准教授 (30260251)
MATSUBARA Hisahiro 千葉大学, 大学院・医学研究院・先端応用外科学, 教授 (20282486)
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| Project Period (FY) |
2008 – 2010
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| Project Status |
Completed (Fiscal Year 2010)
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| Budget Amount *help |
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2010: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2009: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2008: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
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| Keywords | 胃癌 / 化学療法 / 5-FU / チミジル酸合成酵素 / TS-FdUMP複合体 / カルパイン / カルパスタチン / ノックダウン / calpain / calpastatin / protease / p53遺伝子 |
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| Research Abstract |
Thymidylate synthase (TS) plays a major role in the response to 5-fluorouracil (5-FU) by binding directly to the 5-FU metabolite, 5-fluoro-dUMP (FdUMP). The change in the TS expression levels after 5-FU administration was examined in parallel to 5-FU responsiveness in six human gastric adenocarcinoma cell lines to elucidate the source of variability of 5-FU sensitivity. MKN-1, SH-10-TC and MKN-74 cells were more resistant to 5-FU than MKN-28, KATO III and MKN-45 cells. Western-blotting analysis revealed that the 5-FU sensitivity of these cells did not correlate with the basal TS expression levels but did correlate with rapid detection of the TS-FdUMP complex after exposure to 5-FU. In 5-FU-resistant cells, very low levels of the TS-FdUMP complex early after 5-FU exposure were elevated by pretreatment with calpain inhibitors such as benzyloxycarbonyl-leucyl-leucinal (ZLLH) and benzyloxycarbonyl-leucyl-leucyl-leucinal (ZLLLH), but not by other protease inhibitors. In contrastd, ONO-3403, which causes calpain activation, stimulated downregulation of the TS-FdUMP complex in 5-FU-sensitive cells. The expression levels of calpastatin, an endogenous calpain inhibitor, were higher in 5-FU-sensitive cells than in 5-FU-resistant cells. ZLLH increased the 5-FU sensitivity of 5-FU-resistant cells, whereas ONO-3403 decreased the sensitivity of 5-FU-sensitive cells. In addition, knockdown of m-calpain by siRNA increased the 5-FU sensitivity in 5-FU-resistant cells, while knockdown of calpastatin reduced the sensitivity in 5-FU-sensitive cells. These results suggest that calpain might reduce the chemosensitivity of human gastric cancer cells to 5-FU possibly by rapid degradation of the TS-FdUMP complex, a finding that is considered to have novel therapeutic implications.
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Report
(4 results)
Research Products
(11 results)
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[Presentation] Down-regulation of thymidylate synthase-5-fluoro-dUMP complex levels by calpain is implicated in the sensitivity to 5-fluorouracil in human gastric cancer cells.2009
Author(s)
鍋谷圭宏, Suzuki, T., Furuya, A., 瀧口正樹, Ochiai, T., 松原久裕, 日和佐隆樹
Organizer
8th International Gastric Cancer Congress
Place of Presentation
Krakow, Poland
Year and Date
2009-06-11
Related Report
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