| Project/Area Number |
09671218
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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 | Tokyo Medical and Dental University |
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Principal Investigator |
SUGIHARA Kenichi (1998-1999) Tokyo Medical and Dental University, 2nd Dept. of Surg., Professor, 医学部, 教授 (10171167)
東海林 豊 (1997) 東京医科歯科大学, 医学部, 助手 (60251518)
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| Co-Investigator(Kenkyū-buntansha) |
YOSHINAGA Keigo Tokyo Medical and Dental University, 2nd Dept. of Surg., assistant Professor, 医学部, 講師 (80240745)
仁瓶 善郎 東京医科歯科大学, 医学部, 助教授 (00189341)
杉原 健一 東京医科歯科大学, 医学部, 教授 (10171167)
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1999: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1998: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1997: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | LIVER / METASTASIS / CARCINOMA / 大腸癌 / 肝転移 / 肝類洞壁内皮細胞 |
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| Research Abstract |
The liver is the major site for metastasis by colorectal carcinoma (CRC) and may have different mechanisms that inhibit CRC growth. We have used a tumor cell-sinusoidal endothelial cell (SEC) coculture system to evaluate whether SEC are cytotoxic to weakly or highly metastatic CRC cells. SEC were isolated from the livers of normal Swiss mice by a portal vein enzymatic infusion method and established as monolayers in 96 well microtiter plates. Confluent SEC monolayers contained 93% endothelial cells (by low density lipoprotein (LDL) receptor staining) and 7% Kupffer cells. When CRC cells were prelabeled with the vital fluorescent dyes rhodamine-dextran and calcein AM and then cocultured with confluent SEC monolayers to assess viability, the % metabolic activity of Clone A cells, a weakly metastatic CRC, was significantly lower than that of CX-1 cells, a highly metastatic CRC, after 4 hrs of coculture (p<0.05). Pretreatment of SEC gadolinium chloride (GaClィイD23ィエD2), an inhibitor of Kupffer cell function, did not block the effect of SEC on Clone A cell. After 24 hrs of coculture with Clone A, SEC produced 114±5.0 nM nitrite vs 123±5.0 and 8±2.0 for SEC and Clone A cells cultured alone, respectively. Furthermore, when 10ィイD1-6ィエD1 to 1 mM NィイD1GィエD1-methyl-L-arginine (NMMA) was added to SEC and Clone A cocultures, toxicity to Clone A cells was blocked as nitrite production was inhibited by dose greater than 10ィイD1-2ィエD1 mM. Unstimulated murine SEC are more toxic to the weakly metastatic Clone A cells than highly metastatic CX-1 cells. Thus, hepatic SEC may be a major host effector cell population that eliminates weakly metastatic CRC through the production of nitric oxide.
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