Involvement of calcineurin in apoptosis induced by microtubule dysfunction

钙调神经磷酸酶参与微管功能障碍诱导的细胞凋亡

• 批准号: 14580746
• 负责人: YAMAMOTO Hideyuki
• 金额: $ 1.86万
• 依托单位: Kumamoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14580746/
• 关键词: Apoptosis; Bcl-2; Calcineurin; Cancer-cells; Cyclosporin A; Dephosphorylation; FK506; Neurons; 脱燐酸化反応; タキソール

Taxol binds to microtubules and induces apoptosis of various cancer cells. Several lines of evidence indicate that taxol induces apoptosis through activation of c-Jun N-terminal kinase (JNK) and the concomitant phosphorylation of B-cell leukemia/lymphoma-2 (Bcl-2) by the activated JNK. Calcineurin is a serine/threonine protein phosphatase and has been reported to be involved in dephosphorylation of Bcl-2. We found that T24 cells derived from human urinary bladder cancer were resistant to taxol. A combination treatment with taxol and cyclosporin A (CsA) or FK506, calcineurin inhibitors, resulted in loss of calcineurin activity and enhancement of phosphorylation of Bcl-2. In addition, treatment of the cells with CsA or FK506 significantly decreased the expression of Bcl-2 at both the protein and mRNA levels. Interestingly, apoptosis of T24 cells was induced by low of taxol in the presence of CsA or FK506. These results suggest that calcineurin has antiapoptotic concentration of taxol in the presence of CsA or FK506. These results suggest that calcineurin has antiapoptotic actions via the dephosphorylation of Bcl-2 and the induction of the gene expression of Bcl-2. In neurons, tau protein binds to microtubules and regulates the microtubule functions in axons. We found that Ca^<2+>, calmodulin-dependent protein kinase II (CaM kinase II) phosphorylated tau at tubulin binding sites and inhibited the binding of tau to microtubules. These results may suggest that CaM kinase II is involved in apoptosis via the phosphorylation of tau.

紫杉醇与微管结合并诱导各种癌细胞凋亡。多项证据表明，紫杉醇通过激活 c-Jun N 末端激酶 (JNK) 以及激活的 JNK 引起的 B 细胞白血病/淋巴瘤-2 (Bcl-2) 的磷酸化来诱导细胞凋亡。钙调神经磷酸酶是一种丝氨酸/苏氨酸蛋白磷酸酶，据报道参与 Bcl-2 的去磷酸化。我们发现源自人膀胱癌的 T24 细胞对紫杉醇具有耐药性。紫杉醇和环孢菌素 A (CsA) 或 FK506（钙调磷酸酶抑制剂）联合治疗会导致钙调磷酸酶活性丧失并增强 Bcl-2 的磷酸化。此外，用CsA或FK506处理细胞显着降低了Bcl-2在蛋白质和mRNA水平上的表达。有趣的是，在 CsA 或 FK506 存在的情况下，低紫杉醇可诱导 T24 细胞凋亡。这些结果表明，在 CsA 或 FK506 存在的情况下，钙调神经磷酸酶具有抗凋亡浓度的紫杉醇。这些结果表明钙调神经磷酸酶通过 Bcl-2 的去磷酸化和诱导 Bcl-2 的基因表达而具有抗细胞凋亡作用。在神经元中，tau 蛋白与微管结合并调节轴突中的微管功能。我们发现Ca^2+、钙调蛋白依赖性蛋白激酶II(CaM激酶II)在微管蛋白结合位点磷酸化tau并抑制tau与微管的结合。这些结果可能表明 CaM 激酶 II 通过 tau 磷酸化参与细胞凋亡。

项目成果

Yamamoto, H.: "Involvement of Ca^<2+>/calmodulin-dependent phosphorylation of synapsin I in insulin exocytosis."J.Pharmacol.Sci.. 93. 30-34 (2003)

Yamamoto, H.：“胰岛素胞吐作用中突触蛋白 I 的 Ca^2/钙调蛋白依赖性磷酸化的参与。”J.Pharmacol.Sci.. 93. 30-34 (2003)

Nomura, T: "The X-linked inhibitor of apoptosis protein inhibits taxol-induced apoptosis in LNCaP cells."Urological Res.. 31. 37-44 (2003)

Nomura, T：“X 连锁凋亡抑制剂抑制 LNCaP 细胞中紫杉醇诱导的凋亡。”泌尿学研究 31. 37-44 (2003)

Kanasaki, H.: "Regulation of gonadotropin-subunit gene expression by dopamine D_2 receptor agonist in clonal mouse gonadotroph αT3-1 cells."Biol.Reprod.. 67. 1218-1224 (2002)

Kanasaki, H.：“克隆小鼠促性腺激素 αT3-1 细胞中多巴胺 D_2 受体激动剂对促性腺激素亚基基因表达的调节。”Biol.Reprod.. 67. 1218-1224 (2002)

Yonehara, T.: "Characterization of αT3-1 cells stably transfected with luteinizing hormone β-subunit complementary deoxyribonucleic acid."Endocrine J.. 50. 341-354 (2003)

Yonehara, T.：“用黄体生成素 β 亚基互补脱氧核糖核酸稳定转染的 αT3-1 细胞的表征。Endocrine J.. 50. 341-354 (2003)

Kanasaki, H.: "Differential regulation of pituitary hormone secretion and gene expression by thyrotropin-releasing hormone. A role for mitogen-activated protein kinase signaling cascade in rat pituitary GH3 cells."Blol.Reprod.. 67. 107-113 (2002)

Kanasaki, H.：“促甲状腺激素释放激素对垂体激素分泌和基因表达的差异调节。丝裂原激活蛋白激酶信号级联在大鼠垂体 GH3 细胞中的作用。”Blol.Reprod.. 67. 107-113 (2002