Identification of Interferon Responsive Apoptosis Regulating Genes in Hepatocellular Carcinoma

肝细胞癌中干扰素反应性细胞凋亡调节基因的鉴定

• 批准号: 16590632
• 负责人: HIGUCHI Hajime
• 金额: $ 2.24万
• 依托单位: Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16590632/
• 关键词: interferon; hepatocellular carcinoma; cyclooxygenase-2; TRAIL; Death Receptor; apoptosis; caspase; interferon

We identified that cyclooxygenase (COX)-2 gene is markedly upregulated in hepatocellular carcinoma (HCC) cells in response to interferon-beta treatment. Anti-apoptotic function of COX-2 is crucial in HCC cell apoptosis resistance. Apoptosis was identified by 4',6-diamidino-2-phenylindole dihydrochloride (DAPI) and fluorescent staining. IFN-β could up-regulate the expression of TRAIL, while NS-398 increased the expression of TRAIL receptors (especially of death receptor 5). Subsequently, activation of caspase-8 and caspase-3 was observed following the treatment with NS-398 and IFN-β in HepG2 cells. Blockade of TRAIL abrogated cell death induced by a combination of NS-398 and IFN-β in both HepG2 and HuH7 cells. Co-treatment with prostaglandin E2 partially abrogated NS-398+IFN-β-induced cell death in HuH7 cells, but not in HepG2 cells. Furthermore, in vivo experiment showed the combined regimen with NS-398 and IFN-β could reduce the growth of xenotransplated HCCs in nude mice. In conclusion, NS-398 is sufficient to overcome IFN resistance in hepatoma cells through the TRAIL/TRAIL receptor and COX-2 signaling pathway, therefore, the combination would appear to be an new therapeutic regimen for HCC.

我们发现，环氧合酶（考克斯）-2基因在肝细胞癌（HCC）细胞中的干扰素-β治疗的反应显着上调。考克斯-2的抗凋亡作用在肝癌细胞凋亡抵抗中起着重要作用。用4 '，6-二脒基-2-苯基吲哚二盐酸盐（DAPI）和荧光染色法检测细胞凋亡。IFN-β可上调TRAIL的表达，NS-398可上调TRAIL受体（尤其是死亡受体5）的表达。随后，在用NS-398和IFN-β处理HepG 2细胞后，观察到caspase-8和caspase-3的活化。TRAIL的阻断消除了由NS-398和IFN-β的组合在HepG 2和HuH 7细胞中诱导的细胞死亡。与前列腺素E2共处理部分消除了NS-398+IFN-β诱导的HuH 7细胞的细胞死亡，但在HepG 2细胞中没有。体内实验表明，NS-398和IFN-β联合应用能抑制裸鼠肝癌移植瘤的生长。综上所述，NS-398可通过TRAIL/TRAIL受体和考克斯-2信号通路克服肝癌细胞对IFN的耐药性，联合用药有望成为一种新的肝癌治疗方案。

项目成果

Cyclooxygenase (COX)-2 inhibitor and interferon-beta synergistically induce apoptosis in human hepatoma cells in vitro and in vivo

环加氧酶（COX）-2抑制剂和干扰素-β在体内外协同诱导人肝癌细胞凋亡

• 发表时间: 2006
• 期刊: Int-J-Cancer (印刷中)
• 作者: Nobuhiro Nakamoto;Hajime Higuchi;Hideaki Kanamori;Hitomi Horikawa;Satoshi Kurita;Tetsuya Masuda;Shinichiro Tada;Hiromasa Takaishi;Kyoko Toda;Takaya Yamada;Naoki Kumagai;Hidetsugu Saito;Toshifumi Hibi
• 通讯作者: Toshifumi Hibi