Methotrexate significantly induces apoptosis by inhibiting STAT3 activation in NPM-ALK-positive ALCL cells
Methotrexate significantly induces apoptosis by inhibiting STAT3 activation in NPM-ALK-positive ALCL cells
复制标题
甲氨蝶呤通过抑制 NPM-ALK 阳性 ALCL 细胞中 STAT3 的激活显着诱导细胞凋亡
DOI：
10.1016/j.bcp.2019.113666
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发表时间：
2019
影响因子：
5.8
通讯作者：
Funakoshi-Tago Megumi
中科院分区：
文献类型：
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作者：
Uchihara Yuki;Komori Reiko;Tago Kenji;Tamura Hiroomi;Funakoshi-Tago Megumi
间变性大细胞淋巴瘤（ALCL）与特征性染色体易位相关，该易位可产生致癌融合蛋白核磷蛋白-间变性淋巴瘤激酶（NPM-ALK）。甲氨蝶呤是一种常用的化疗药物，由于其抑制二氢叶酸还原酶（DHFR），抑制DNA的合成，在治疗多种癌症。在本研究中，我们发现低剂量甲氨蝶呤通过抑制NPM-ALK关键下游分子信号转导和转录因子激活因子3（STAT 3）的激活，显著诱导表达NPM-ALK的转化Ba/F3细胞凋亡。虽然甲氨蝶呤阻止了STAT 3的磷酸化，但它不影响NPM-ALK的活性。与亚叶酸的共同处理阻止了甲氨蝶呤诱导的STAT 3激活抑制和细胞凋亡诱导，表明甲氨蝶呤通过NPM-ALK消耗转化细胞中的四氢叶酸（THF）发挥其细胞毒性作用。此外，甲氨蝶呤诱导抗凋亡蛋白MCL-1的下调、DNA损伤和p53肿瘤抑制因子的激活，通过抑制STAT 3导致细胞凋亡。甲氨蝶呤显著诱导了表达携带点突变G262 R的NPM-ALK突变体的ALK耐药细胞和ALCL患者源性NPM-ALK阳性Ki-JK细胞的凋亡。总之，这些结果证明了甲氨蝶呤（抑制STAT 3活化）对NPM-ALK阳性ALCL的潜在治疗应用。
Anaplastic large cell lymphoma (ALCL) is associated with a characteristic chromosomal translocation that generates the oncogenic fusion protein, nucleophosmin-anaplastic lymphoma kinase (NPM-ALK). Methotrexate is a commonly used chemotherapeutic drug in the treatment of multiple cancers due to its inhibition of dihydrofolate reductase (DHFR), which suppresses the synthesis of DNA. In the present study, we found that low-dose methotrexate significantly induced apoptosis in transformed Ba/F3 cells expressing NPM-ALK by inhibiting the activation of signal transducer and activator of transcription factor 3 (STAT3), a critical downstream molecule of NPM-ALK. Although methotrexate prevented the phosphorylation of STAT3, it did not affect the activity of NPM-ALK. A co-treatment with folinic acid prevented the methotrexate-induced inhibition of STAT3 activation and induction of apoptosis, suggesting that methotrexate exerts its cytotoxic effects by depleting tetrahydrofolate (THF) in transformed cells by NPM-ALK. Furthermore, methotrexate induced the down-regulation of the anti-apoptotic protein, MCL-1, DNA damage, and the activation of a p53 tumor suppressor, leading to apoptosis through the inhibition of STAT3. Methotrexate significantly induced apoptosis in ALK inhibitor-resistant cells expressing the NPM-ALK mutant harboring the point mutation, G262R, and in ALCL patient-derived NPM-ALK-positive Ki-JK cells. Collectively, these results demonstrate the potential therapeutic application of methotrexate, which inhibits the activation of STAT3, to NPM-ALK-positive ALCL.