| Project/Area Number |
16590614
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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 | Nagoya City University |
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Principal Investigator |
JOH Takashi Nagoya City University, Graduate School of Medical, Sciences, Professor, 大学院・医学研究科, 教授 (30231369)
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| Co-Investigator(Kenkyū-buntansha) |
HIGASHIYAMA Shigeki Ehime University, School of Medical Sciences, Professor, 医学部, 教授 (60202272)
OHARA Hirotaka Nagoya City University, Graduate School of Medical, Sciences, Assistant Professor, 大学院・医学研究科, 講師 (80285212)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2005: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2004: ¥2,600,000 (Direct Cost: ¥2,600,000)
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| Keywords | EGF / HB-EGF C terminus / ADAM / trafficking into nucleu / HB-EGF C 末端 / HB-EGF / PLZA / c-myc / C末端 |
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| Research Abstract |
We have recently shown that Helicobacter pylori as a causal of gastric cancer induces IL-8 production from gastric epithelia, IL-8 subsequently promotes cell migration and proliferation through metalloproteinase-cleavage proHB-EGF. HB-EGF C terminus, trafficking into nucleus, subsequently exerts some effects on regulation of cell growth in fibroblastoma cells. However, little is known about whether this mechanism exists in gastric epithelia. Thus, the presence of HB-EGF C terminus trafficking into nucleus was examined with gastric epithelia and gastric cancer tissues.
(method)Gastric cell lines(KATO III, GCIY)were treated with a 6OnM of phorbor ester with or without ADAM inhibitor(KB-R7785). Cell lysates were probed with antibodies against phosphorylation after immunoprecipitation with anti-EGFR antibodies. Cells were transfected with vectors expressing YFP fused to C terminus of proHB-EGF. Cells were treated with a 60nM of phorbor ester with or without KB-R7785, and then were observed under a fluorescent microscopy. Excided stomach samples were immunohistochemically stained with anti-bodies against HB-EGF C-terminus.
(result)In KATO III cells, EGFR phosphorylation by phorbor ester peaked within 15 min. That phosphorylation was abolished by KB-R7785. HB-EGF C terminus trafficked into nucleus from membrane with time. In KATO III and GCIY cells, HB-EGF C terminus was stained in the nuclei 60min after stimulation. In carcinomatous components of excided advanced gastric cancer, HB-EGF C terminus was partially stained in the nuclei.
(conclusion)ADAM plays an important roles in the EGF signaling pathways, where ADAM cleaves extracellular domain of proHB-EGF and C terminus of proHB-EGF trafficks into the nucleus in gastric cancer cells. Thus, regulation of ADAM may be helpful for control of cell growth, migration and cell cycle, and leads to possible strategy against gastric cancer.
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