Analysis of signal transduction in human glioma cells transfected with cytokine gene

转染细胞因子基因的人胶质瘤细胞信号转导分析

• 批准号: 07457312
• 负责人: YOSHIDA Jun
• 金额: $ 4.8万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07457312/
• 关键词: BRAIN TUMOR; INTERFERON; TNF; GENE; LIPOSOME; NF-kappaB; PHOSPHORYLATION; SIGNAL TRANSDUCTION; NF-κB

Our aim is analysis of mechanisms of antitumor activity induced by cytokine gene transfection into human glioma cells.To clarify the underlying mechanisms involved in the different susceptibility to cytokines, we examined the degrees of phosphorylation or NF-kappaB complex when cytokines (interferon-beta (IFN-beta) or tumor necrosis factor-alpha (TNF-alpha)) were exogenously added to human glioma cells. These results suggest that the different NF-kB complex induced by cytokines in human glioma cells might be related to the varying degrees of susceptibility of tumor cells to cytokines.To elucidate the mechanisms underlying the antitumor activity of cytokine gene transfection using cationic liposomes, we observed morphological changes under video-enhanced contrast differential interference contrast microscopy. Cells harboring the intranuclear human IFN-beta gene underwent morphological changes characterized by bled formation and cytoplasmic boiling such as apoptosis. On the other hand, exogenously added cytokines did not induce the changes at all.In in vivo experiments using C57BL/6 mouse, we confirmed the activation of cellular immunity when liposomes, containing mouse IFN-beta gene were injected into the experimental glioma.

我们的目的是分析细胞因子基因转染人胶质瘤细胞诱导抗肿瘤活性的机制。为了阐明不同细胞因子敏感性的潜在机制，我们检测了细胞因子(干扰素- β （ifn - β)或肿瘤坏死因子- α (tnf - α)）外源性添加到人类胶质瘤细胞时磷酸化或nf - κ b复合物的程度。这些结果提示，细胞因子在人胶质瘤细胞中诱导的不同NF-kB复合物可能与肿瘤细胞对细胞因子的不同易感性程度有关。为了阐明阳离子脂质体转染细胞因子基因抗肿瘤活性的机制，我们在视频增强对比差干涉对比显微镜下观察了形态学变化。核内携带人类ifn - β基因的细胞发生了以出血形成和细胞质沸腾为特征的形态学变化，如凋亡。另一方面，外源添加的细胞因子完全没有引起这些变化。在C57BL/6小鼠的体内实验中，我们证实了将含有小鼠ifn - β基因的脂质体注射到实验性胶质瘤中可以激活细胞免疫。

项目成果

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