The PI3K/Akt/GSK-3β/ROS/eIF2B pathway promotes breast cancer growth and metastasis via suppression of NK cell cytotoxicity and tumor cell susceptibility
The PI3K/Akt/GSK-3β/ROS/eIF2B pathway promotes breast cancer growth and metastasis via suppression of NK cell cytotoxicity and tumor cell susceptibility
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PI3K/Akt/GSK-3 beta/ROS/eIF2B 通路通过抑制 NK 细胞的细胞毒性和肿瘤细胞易感性促进乳腺癌生长和转移
DOI：
10.20892/j.issn.2095-3941.2018.0253
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发表时间：
2019-02-01
影响因子：
5.5
通讯作者：
Wang, Yue
中科院分区：
文献类型：
--
作者：
Jin, Fengjiao;Wu, Zhaozhen;Wang, Yue
目的：为了检测pSer 9-GSK-3 β对乳腺癌的作用，并确定潜在的代谢和免疫机制是否与ROS/eIF 2B和自然杀伤（NK）细胞有关，我们使用TWS 119通过磷酸化Ser 9来修饰GSK-3 β，并探讨其对乳腺癌和NK细胞的作用。通过PCR和Western印迹定量GSK-3 β、自然杀伤组2成员D（NKG 2D）配体、eIF 2B的表达。我们分别用DCFH-DA和MitoSOX（TM）探针测定细胞内活性氧（ROS）和线粒体ROS，并用线粒体呼吸链复合物I/III试剂盒对4 T1细胞进行细胞呼吸定量分析。我们的研究显示TWS 119下调NKG 2D配体，（H60 a和Rae 1），抑制NK细胞的细胞毒性，并促进4 T1小鼠乳腺癌细胞的迁移。然而，LY 290042通过抑制PI 3 K/Akt通路来减弱p-GSK-3 β的形成，逆转了这些作用。我们还发现，pSer 9-GSK-3 β的高表达诱导了较高水平的ROS，并观察到线粒体呼吸链复合物I/III功能的异常诱导GSK-3 β诱导的电子传递链功能障碍，从而自然干扰ROS水平。另外，乳腺癌组织中NOX 3和NOX 4的表达显著上调，影响ROS的产生，与乳腺癌的转移有关。结论：PI 3 K/Akt/GSK-3 β/ROS/eIF 2B信号通路可调节NK细胞活性和肿瘤细胞对NK细胞的敏感性，从而导致乳腺癌的生长和肺转移。因此，GSK-3 β是一个有前途的抗肿瘤治疗靶点。
Objective: To examine the effect of pSer9-GSK-3 beta on breast cancer and to determine whether the underlying metabolic and immunological mechanism is associated with ROS/eIF2B and natural killer (NK) cells.Methods: We employed TWS119 to inactivate GSK-3 beta by phosphorylating Ser9 and explored its effect on breast cancer and NK cells. The expression of GSK-3 beta, natural killer group 2 member D (NKG2D) ligands, eIF2B was quantified by PCR and Western blot. We measured intracellular reactive oxygen species (ROS) and mitochondrial ROS using DCFH-DA and MitoSOX (TM) probe, respectively, and conducted quantitative analysis of cellular respiration on 4T1 cells with mitochondrial respiratory chain complex I/III kits.Results: Our investigation revealed that TWS119 downregulated NKG2D ligands (H60a and Rae1), suppressed the cytotoxicity of NK cells, and promoted the migration of 4T1 murine breast cancer cells. Nevertheless, LY290042, which attenuates p-GSK-3 beta formation by inhibiting the PI3K/Akt pathway, reversed these effects. We also found that higher expression of pSer9-GSK-3 beta induced higher levels of ROS, and observed that abnormality of mitochondrial respiratory chain complex I/III function induced the dysfunction of GSK-3 beta-induced electron transport chain, naturally disturbing the ROS level. In addition, the expression of NOX3 and NOX4 was significantly up-regulated, which affected the generation of ROS and associated with the metastasis of breast cancer. Furthermore, we found that the expression of pSer535-eIF2B promoted the expression of NKG2D ligands (Mult-1 and Rae1) following by expression of pSer9-GSK-3 beta and generation of ROS.Conclusions: The PI3K/Akt/GSK-3 beta/ROS/eIF2B pathway could regulate NK cell activity and sensitivity of tumor cells to NK cells, which resulted in breast cancer growth and lung metastasis. Thus, GSK-3 beta is a promising target of anti-tumor therapy.