| Project/Area Number |
21590421
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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 |
Experimental pathology
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| Research Institution | Keio University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
OKADA Yasunori 慶應義塾大学, 医学部, 教授 (00115221)
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| Project Period (FY) |
2009 – 2011
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| Project Status |
Completed (Fiscal Year 2011)
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| Budget Amount *help |
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2011: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2010: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2009: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
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| Keywords | ADAM / 癌転移 / VWF / アポトーシス / MMP / CTGF / 血管新生 / 乳癌 / バイオイメージング / 肺癌 / 転移 / 浸潤 / ルシフェラーゼ |
|---|
| Research Abstract |
To explore the molecular mechanism of ADAM28/VWF-mediated metastasis, we established PC-9 lung carcinoma cells and MDA-MB231 breast carcinoma cells expressing Venus and luciferase(PC-9^<ffLuc-cp156> and MDA-MB231^<ffLuc-cp156> cells), and used them in the mouse models of lung metastasis, and primary tumor growth and spontaneous metastasis. Inhibition of ADAM28 expression with shRNA or siRNA and its inactivation with the neutralizing anti-ADAM28 antibody significantly reduced lung metastasis of PC-9^<ffLuc-cp156> cells and enhanced apoptosis of the cells within blood vessels. The primary tumor growth of ADAM28-shRNA transfectants after orthotopic implantation of MDA-MB231^<ffLuc-cp156> cells was significantly slower than that of mock-transfectants. By RT-PCR analysis for the expression of ffLuc-cp156 transcripts, metastasis in the lung, heart, liver, kidney and brain was reduced in the mice received ADAM28-shRNA transfectants as compared with mock-transfectants. Plasma samples from mice with injection of mock-transfected PC-9^<ffLuc-cp156> cells showed decreased amount of large VWF multimers as compared with those from mice which received ADAM28-shRNA transfectants or no cells. All these in vivo and in vitro data suggest that ADAM28 derived from carcinoma cells cleaves and inactivates pro-apoptotic endogenous agent VWF, thereby promotes the carcinoma cell survival within the blood stream and ultimately enhances their metastasis
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