The mechanism of apoptosis induction by TGF-β stimuli and its clinical application

TGF-β刺激诱导细胞凋亡的机制及其临床应用

• 批准号: 13670157
• 负责人: OSADA Hirotaka
• 金额: $ 2.24万
• 依托单位: Aichi Cancer Center
• 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-13670157/
• 关键词: hepatoma; TGF-β; Apoptosis; 肝癌

We studied the mechanism of apoptosis induction by TGF-β stimuli for the purpose of applying in the cancer therapy. We established several apoptosis-inducible and -uninducible subclones derived from the single hepatoma cell line, which were partially sensitive to TGF-β stimuli. Our preliminary study using apoptosis-specific cDNA arrays showed that the expression of TNF family members were induced by TGF-β stimuli in the apoptosis-inducible subclones. We studied further the signaling pathways from TGF-β stimuli to apoptosis. After TGF-β stimuli, DNA contents, expression of TNF family, and activation of caspase family were sequentially studied. At first, the G1/G2 arrest was transiently observed, and then the induction of TNF family expression, caspase-8 activation, and apoptosis induction occurred sequentially. We studied the effects of neutralizing antibodies against TNF family members and caspase-8 inhibitor, Z-IETD-FMK against the apoptosis induction, and obtained about 50% inhibition of apoptosis. The NF- κB reporter analysis also showed the activation of NF-κB pathway by TGF-β stimuli. Our studies demonstrated the signaling pathway from TGF-β to apoptosis through TNF family induction and caspase-8 activation. Furhter studies using high-density cDNA array may be required to reveal the whole pathways of apoptosis induction. The activation of NF-κB might be associated with the TGF-β -resistance, but it remains to be determined.

我们研究了TGF-β刺激诱导细胞凋亡的机制，以期应用于癌症治疗。我们从单个肝癌细胞系中建立了几个对TGF-β刺激部分敏感的凋亡诱导型和非诱导型亚克隆。我们利用肿瘤特异性cDNA阵列的初步研究表明，在肿瘤诱导亚克隆中，TNF家族成员的表达受到TGF-β刺激的诱导。我们进一步研究了从TGF-β刺激到细胞凋亡的信号通路。在TGF-β刺激后，依次观察DNA含量、TNF家族表达和caspase家族活化。首先观察到短暂的G1/G2期阻滞，然后依次诱导TNF家族表达、caspase-8激活和凋亡诱导。我们研究了TNF家族成员中和抗体和caspase-8抑制剂Z-IETD-FMK对细胞凋亡诱导的影响，获得了约50%的细胞凋亡抑制。NF- κB报告基因分析也显示TGF-β刺激激活了NF-κB通路。我们的研究证实了TGF-β通过TNF家族的诱导和caspase-8的激活而导致细胞凋亡的信号通路。利用高密度cDNA芯片的进一步研究可能需要揭示细胞凋亡诱导的整个途径。NF-κB的活化可能与TGF-β抵抗有关，但其作用机制尚不明确。

项目成果

Osada H, Tatematsu Y, et al.: "Frequent and histological type-specific inactivation of 14-3-3s in human lung cancers"Oncogene. 21. 2418-2424 (2002)

Osada H、Tatematsu Y 等人：“人类肺癌中 14-3-3 的频繁和组织学类型特异性失活”癌基因。

Osada H, Tatematsu Y, et al.: "Heterogeneous transforming growth factor (TGF)-b unresponsiveness and loss of TGF-b receptor type II expression caused by histone deacetylation in lung cancer cell lines"Cancer Research. 61. 8331-8339 (2001)

Osada H、Tatematsu Y 等人：“肺癌细胞系中组蛋白脱乙酰化引起的异质转化生长因子 (TGF)-b 无反应和 TGF-b 受体 II 型表达缺失”癌症研究。

Mizuno K, Osada H, Konishi H, Tatematsu Y, Yatabe Y, Mitsudomi T, Fujii Y, and Takahashi T: "Aberrant hypermethylation of the CHFR prophase checkpoint gene in human lung cancers"Oncogene. 21. 2328-2333 (2002)

Mizuno K、Osada H、Konishi H、Tatematsu Y、Yatabe Y、Mitsudomi T、Fujii Y 和 Takahashi T：“人类肺癌中 CHFR 前期检查点基因的异常高甲基化”癌基因。

Konishi H, Osada H, et al.: "Identification of frequent G(2) checkpoint impairment and a homozygous deletion of 14-3-3ε At 17p13.3 in small cell lung cancers"Cancer Research. 62. 271-276 (2002)

Konishi H、Osada H 等人：“小细胞肺癌中频繁 G(2) 检查点损伤和 1​​4-3-3ε At 17p13.3 纯合缺失的鉴定”癌症研究。 62. 271-276 (2002) ）

Konishi, H., Sugiyama, M., Mizuno, K., Saito, H., Yatabe, Y., Takahashi, T., Osada. H., and Takahashi T: "Detailed characterization of a homozygously deleted region corresponding to a candidate tumor suppressor locus at distal 17p13.3 in human lung cancer

小西，H.，杉山，M.，水野，K.，斋藤，H.，矢田部，Y.，高桥，T.，长田。