Gene therapy against hepatocellular carcinoma using p48 gene

使用p48基因针对肝细胞癌的基因治疗

• 批准号: 13670529
• 负责人: EJIMA Eri
• 金额: $ 2.18万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670529/
• 关键词: p48; Interferon-α; eIF2 α; PKR; Hepatoma; TRAIL; IFN-α; eIF2α

Double-stranded RNA-dependent protein kinase (PKR) is a key factor involved in interferon (IFN)-induced antiviral actions. Since p48, together with signal transducers and activators of transcription 1 and 2 (STAT1 and STAT2), is an indispensable mediator in IFN-α signaling pathways, we investigated the effect of p48 gene transduction on PKR expression and its activity in HuH-7 human hepatoma cells.HuH-7 cells were infected or transfected with p48 gene expression adenoviral vector or plasmid vector, respectively, and incubated with or without IFN-α, then PKR expression and phosphorylation of α-subunit of eukaryotic protein synthesis initiation factor-2 (eIF2 α) in the cells were examined. In addition, PKR activity inhibiting protein translation was determined by the decrease of chloramphenicol acetyltransferase (CAT) gene translation or α-fetoprotein secretion.p48 overexpression itself could not stimulate PKR expression. However, p48 overexpression in combination with interferon-α treatment caused a marked increase in PKR expression and augmented the phosphorylation of eIF2 α, by which the transfected CAT gene translation, as well as the endogenous α-fetoprotein synthesis, was blocked without affecting their mRNA levels.These results suggest that p48 gene transduction may provide a strategy to enhance the IFN-mediated PKR expression and its activity in hepatocytes.

双链RNA依赖性蛋白激酶（PKR）是干扰素（IFN）诱导抗病毒作用的关键因子。由于p48，连同信号转导和转录激活因子1和2为了研究p48基因转导对人肝癌细胞HuH-7 PKR表达及其活性的影响，分别用p48基因表达腺病毒载体和质粒载体感染HuH-7细胞，分别与IFN-α共同孵育，检测PKR的表达及eIF 2 α亚基的磷酸化水平。此外，PKR抑制蛋白翻译的活性取决于氯霉素乙酰转移酶（CAT）基因翻译或甲胎蛋白分泌的减少，p48过表达本身并不能刺激PKR的表达。然而，p48过表达与干扰素-α联合治疗引起PKR表达显著增加，并增加eIF 2 α的磷酸化，从而使转染的CAT基因翻译以及内源性甲胎蛋白的合成，这些结果表明，p48基因转导可能提供了一种增强IFN-γ表达的策略。介导的PKR表达及其在肝细胞中的活性。

项目成果

Yoko Tamada, et al.: "p48 overexpression enhances interferon-mediated expression and activity of double-stranded RNA-dependent protein kinase (PKR) in human hepatoma cells"Journal of Hepatology. 37・4. 493-499 (2002)

Yoko Tamada 等人：“p48 过度表达增强人肝癌细胞中干扰素介导的双链 RNA 依赖性蛋白激酶 (PKR) 的表达和活性”《肝脏病学杂志》37·4 (2002)。

Yoko Tamada, et al.: "p48 overexpression enhances interferon-mediated expression and activity of double-stranded RNA-dependent protein kinase in human hepatoma cells"Journal of Hepatology. 37. 493-499 (2002)

Yoko Tamada 等人：“p48 过表达增强人肝癌细胞中干扰素介导的双链 RNA 依赖性蛋白激酶的表达和活性”《肝脏病学杂志》。

Yoko Tamada, Kazuhiko Nakao, Yuji Nagayama, Keisuke Nakata, Tatsuki Ichikawa, Yosei Kawamata, Hiroki Ishikawa, Keisuke Hamasaki, Katsumi Eguchi, Nobuko Ishii: "p48 overexpression enhances interferon-medialed expression and activity of double-stranded RNA-

Yoko Tamada、Kazuhiko Nakao、Yuji Nagayama、Keisuke Nakata、Tatsuki Ichikawa、Yosei Kawamata、Hiroki Ishikawa、Keisuke Hamasaki、Katsumi Eguchi、Nobuko Ishii：“p48 过表达增强了干扰素介导的双链 RNA 的表达和活性

Masaya Shigeno, et al.: "Interferon-α sensitizes human hepatoma cells to TRAIL-induced apoptosis through DR5 upregulation and NF-κB inactivation"Oncogene. (in press). (2003)

Masaya Shigeno 等人：“干扰素-α 通过 DR5 上调和 NF-κB 失活使人肝癌细胞对 TRAIL 诱导的细胞凋亡敏感”Oncogene（出版中）。

Masaya Shigeno, et al.: "Interferon-α sensitizes human hepatoma cells to TRAIL-induced apoptosis through DR5 upregulation and NF-κB inactivation"Oncogene. 22. 1653-1662 (2003)

Masaya Shigeno 等人：“干扰素-α 通过 DR5 上调和 NF-κB 失活使人肝癌细胞对 TRAIL 诱导的细胞凋亡敏感”Oncogene。22. 1653-1662 (2003)