ENDOTHELIN-1 IS AN AUTOCRINE GROWTH FACTOR FOR HEPATOCELLULAR CARCINO

内皮素-1 是肝细胞癌的自分泌生长因子

• 批准号: 05454324
• 负责人: ISHIBASHI Miyuki
• 金额: $ 3.9万
• 依托单位: Teikyo University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05454324/
• 关键词: endothelin; growth factor; hepatocellular carcinoma; tomor growth; endothelin receptor; autocrine growth factor; receptor antagonist

To test the hypothesis that endothelin-1 (ET-1) may be an autocrine or paracrine growth factor for hepatocellular carcinoma (HCC), we examined the in vitro production and secretion of ET-1 by human HCC cell lines and ET-1 receptors on those cells, and studied the effect of ET-1 and/or specific ET receptor antagonists on HCC growth. Northern blot analysis demonstrated the expression of mRNA for ET-1 precursor in HCC cell lines and a substantial amount of ET-1 as determined by a specific enzyme immunoassay was found both in cell extracts and in culture media. Immunohistochemistry of HCC cells showed HCC cells positive for ET-1. When HCC cells were grown in semi-solid medium containing ET-1, ET-1 markedly increased colony growth of these HCC cell lines in a dose-dependent manner. When 10 ^6M BQ-123, a specific ET receptor antagonist, was coincubated, the stimulatory effect of 10 ^8M ET-1 on the growth of HCC cells was completely blocked. Furthermore, colony formation of Mahlavu and Alexan … More der cells was significantly inhibited by incubation with BQ-123 or the monoclonal antibody for ET-1 alone. Northern blot analysis demonstrated the expression of mRNA for human ET_B receptor in at least two HCC cell lines, Mahlavu and Hep G2. The ET_A receptor mRNA was not expressed in any of the HCC cell lines examined. A binding study conducted using a labeled endothelin as a ligand showed high affinity binding sites on Huh-7 cells (Kd=2.7 x 10 ^<10>M). To determine whether HCC growth may be ET-dependent then, two HCC cell lines, Mahlavu and Huh-7, were implanted into athymic nude mice. Administration of bosentan (10 mg/kg/day) caused a significant inhibition of tumor growth in both experiments. These results indicate that ET-1 is produced and secreted by HCC,which stimulates its own growth, and that HCC cells are enriched with ET_B receptors, which may mediate the growth-stimulatory effect of ET-1 on HCC.We conclude that ET-1 is a growth factor for HCC,and that ET receptor antagonists may be useful in the medical treatment of HCC. Less

为了验证内皮素-1 （ET-1）可能是肝细胞癌（HCC）的自分泌或旁分泌生长因子的假设，我们检测了人肝癌细胞系和ET-1受体在体外产生和分泌ET-1，并研究了ET-1和/或特异性ET受体拮抗剂对HCC生长的影响。Northern blot分析表明，在HCC细胞系中有ET-1前体mRNA的表达，并且在细胞提取物和培养基中都发现了大量的ET-1，这是通过特定的酶免疫测定法确定的。肝癌细胞免疫组化显示ET-1阳性。当HCC细胞在含有ET-1的半固体培养基中生长时，ET-1显著增加这些HCC细胞系的集落生长，并呈剂量依赖性。当特异性ET受体拮抗剂10 ^6M BQ-123共孵育时，10 ^8M ET-1对HCC细胞生长的刺激作用被完全阻断。此外，BQ-123或ET-1单克隆抗体孵育可显著抑制Mahlavu和Alexan…More der细胞的集落形成。Northern blot分析显示，人类ET_B受体mRNA在至少两种HCC细胞系Mahlavu和Hep G2中表达。ET_A受体mRNA在所有HCC细胞系中均未表达。使用标记内皮素作为配体进行的结合研究显示，Huh-7细胞具有高亲和力的结合位点（Kd=2.7 x 10 ^<10>M）。为了确定肝细胞癌的生长是否依赖et，我们将两种肝细胞癌细胞系Mahlavu和Huh-7植入胸腺裸鼠体内。在两个实验中，波生坦（10mg /kg/天）对肿瘤生长均有显著抑制作用。这些结果表明，ET-1是由HCC产生和分泌的，能够刺激其自身的生长，而HCC细胞中富含ET_B受体，可能介导ET-1对HCC的促生长作用。我们得出结论，ET-1是HCC的生长因子，ET受体拮抗剂可能在HCC的医学治疗中有用。少

项目成果

Kamoi K,Sudo N,Ishibashi M,Yamaji T: "Plasma endothelin-1 levels in patients with preqnancy-induced hypertension." N Engl J Med. 323. 1486-1487 (1990)

Kamoi K、Sudo N、Ishibashi M、Yamaji T：“妊娠期高血压患者的血浆内皮素-1 水平。”

Sudo N,Kamoi K,Ishibashi M,Yamaji T: "Plasma endothelin-1 and big endothelin-1 levels in women with pre-eclampsia." Acta Endocrinol. 129. 114-120 (1993)

Sudo N、Kamoi K、Ishibashi M、Yamaji T：“先兆子痫女性的血浆内皮素 1 和大内皮素 1 水平。”

Ishibashi M,Fujisawa M,Furue H,Maeda Y,Fukayama M,Yamaji T: "Inhibition of growth of human small cell lung cancer by bromocriptine." Cancer Res. 54. 3442-3446 (1994)

Ishibashi M、Fujisawa M、Furue H、Maeda Y、Fukayama M、Yamaji T：“溴隐亭抑制人小细胞肺癌的生长。”

Ishibashi M,Fujita M,Nagai K,Kako K,Furue H,Kaku E,Osamura Y,Yamaji T: "Production and secretion of endothelin by hepatocellular carcinoma." J Clin Endocrinol Metab. 76. 378-383 (1993)

Ishibashi M，Fujita M，Nagai K，Kako K，Furue H，Kaku E，Osamura Y，Yamaji T：“肝细胞癌内皮素的产生和分泌。”

石橋みゆき,古江尚,大西真,三浦英明,藤沢道夫,岡輝明,山路徹: "成長因子としてのエンドセリン." 成長科学協会 研究年報 平成5年度. 17. 343-354 (1994)

Miyuki Ishibashi、Hisashi Furue、Makoto Onishi、Hideaki Miura、Michio Fujisawa、Teruaki Oka、Toru Yamaji：“内皮素作为生长因子”1993. 17. 343-354 (1994)