Metformin induces apoptotic cytotoxicity depending on AMPK/PKA/GSK-3β-mediated c-FLIPL degradation in non-small cell lung cancer
Metformin induces apoptotic cytotoxicity depending on AMPK/PKA/GSK-3β-mediated c-FLIPL degradation in non-small cell lung cancer
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DOI：
10.2147/cmar.s178688
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发表时间：
2019-01-01
影响因子：
3.3
通讯作者：
Shi, Shaoqing
中科院分区：
文献类型：
--
作者：
Luo, Zhuang;Zhu, Tingting;Shi, Shaoqing
背景资料：二甲双胍是一种一线抗糖尿病药物，最近报道其在多种癌症中具有抗癌活性;然而，其潜在机制仍然难以捉摸。本研究的目的是探讨细胞FADD样IL-1 β转化酶（FLICE）抑制蛋白大（c-FLIPL）在二甲双胍诱导的非小细胞肺癌（NSCLC）体外抗肿瘤活性中的作用。实时荧光定量PCR检测相关基因mRNA水平。Western blot检测相关蛋白的表达。结果：美托洛尔能明显抑制A549和H460细胞的增殖，并呈剂量依赖性。机制研究表明，二甲双胍通过诱导凋亡性细胞死亡杀死NSCLC细胞。此外，二甲双胍大大抑制c-FLIPL表达，然后促进其降解。此外，二甲双胍显著激活腺苷5 '-单磷酸（AMP）活化蛋白激酶（AMPK）及其下游糖原合成酶激酶3 β（GSK-3 β），阻断AMPK的表达，而GSK-3 β与siRNA部分逆转二甲双胍诱导的细胞毒性，并恢复肺癌细胞中c-FLIPL的表达。结论：二甲双胍通过AMPK/PKA/GSK-3 β轴介导的c-FLIPL降解作用杀伤NSCLC细胞。
Background: Metformin, a first-line antidiabetic drug, has recently been reported with anticancer activities in various cancers; however, the underlying mechanisms remain elusive. The aim of the present study was to investigate the role of cellular FADD-like IL-1 beta-converting enzyme (FLICE)-inhibitory protein large (c-FLIPL) in metformin-induced anticancer activity in non-small cell lung cancer (NSCLC) in vitro.Materials and methods: Cell viability was measured by MTT assay. Quantitative real-time PCR was carried out to detect the level of mRNA of related genes. The expression of related proteins was detected by Western blot. siRNA was used to silence the expression of targeted proteins.Results: Metformin significantly suppressed proliferation of both A549 and H460 cells in a dose-dependent manner. Mechanistic studies suggested that metformin killed NSCLC cells by inducing apoptotic cell death. Moreover, metformin greatly inhibited c-FLIPL expression and then promoted its degradation. Furthermore, metformin significantly activated Adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) and its downstream glycogen synthase kinase 3beta (GSK-3 beta), block the expression of AMPK, and GSK-3 beta with siRNA partially reversed metformin-induced cytotoxicity and restored the expression of c-FLIPL in lung cancer cells. Metformin also suppressed the activity of AMPK downstream protein kinase A (PKA), PKA activators, both 8-Br-cAMP and forskolin, greatly increased c-FLIPL expression in NSCLC cells.Conclusion: This study provided evidence that metformin killed NSCLC cells through AMPK/PKA/GSK-3 beta axis-mediated c-FLIPL degradation.