Probucol suppresses human glioma cell proliferation in vitro via ROS production and LKB1-AMPK activation (Retracted article. See vol. 39, pg. 328, 2018)
Probucol suppresses human glioma cell proliferation in vitro via ROS production and LKB1-AMPK activation (Retracted article. See vol. 39, pg. 328, 2018)
复制标题
普罗布考通过 ROS 产生和 LKB1-AMPK 激活抑制体外人胶质瘤细胞增殖
DOI：
10.1038/aps.2014.88
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发表时间：
2014-12-01
影响因子：
8.2
通讯作者：
Wang, Fu-rong
中科院分区：
文献类型：
--
作者：
Jiang, Yong-sheng;Lei, Jing-an;Wang, Fu-rong
目的：普罗布考是一种抗高血压药物，在肿瘤发生、发展和进展的各个阶段均具有抗肿瘤活性。本研究旨在探讨该药是否影响体外培养的胶质瘤细胞的生长及其作用机制。使用细胞增殖测定和BrdU掺入来评估细胞增殖。Western blot检测AMPK、肝激酶B1（LKB 1）和p27（Kip 1）的磷酸化。26 S蛋白酶体活性用原位荧光底物法测定。结果：普罗布考（10-100 μ mol/L）处理U87胶质瘤细胞后，细胞增殖受到抑制，并呈剂量和时间依赖性。同时，普罗布考可显著增加细胞内活性氧的产生，使AMPK的Thr 172和LKB 1的Ser 428磷酸化。此外，普罗布考显着降低26 S蛋白酶体活性和增加p27（Kip 1）蛋白水平在细胞中的AMPK依赖性的方式。用tempol（一种超氧化物歧化酶模拟物）、MG 132（蛋白酶体抑制剂）或化合物C（AMPK抑制剂）预处理或通过LKB 1、AMPK或p27（Kip 1）基因沉默可逆转普罗布考诱导的U87细胞增殖抑制。结论：Probucol通过产生ROS和激活LKB 1-AMPK，减少26 S蛋白酶体依赖的p27（Kip 1）降解，抑制人胶质瘤细胞的增殖。
Aim: Probucol, an anti-hyperlipidemic drug, has been reported to exert antitumor activities at various stages of tumor initiation, promotion and progression. In this study we examined whether the drug affected glioma cell growth in vitro and the underlying mechanisms.Methods: Human glioma U87 and glioblastoma SF295 cell lines were used. Cell proliferation was accessed using the cell proliferation assay and BrdU incorporation. The phosphorylation of AMPK, liver kinase B1 (LKB1) and p27(Kip1) was detected by Western blot. The activity of 26S proteasome was assessed with an in situ fluorescent substrate. siRNAs were used to suppress the expression of the relevant signaling proteins.Results: Treatment of U87 glioma cells with probucol (10-100 mu mol/L) suppressed the cell proliferation in dose-and time-dependent manners. Meanwhile, probucol markedly increased the ROS production, phosphorylation of AMPK at Thr172 and LKB1 at Ser428 in the cells. Furthermore, probucol significantly decreased 26S proteasome activity and increased p27(Kip1) protein level in the cells in an AMPK-dependent manner. Probucol-induced suppression of U87 cell proliferation could be reversed by pretreatment with tempol (a superoxide dismutase mimetic), MG132 (proteasome inhibitor) or compound C (AMPK inhibitor), or by gene silencing of LKB1, AMPK or p27(Kip1). Similar results were observed in probucol-treated SF295 cells.Conclusion: Probucol suppresses human glioma cell proliferation in vitro via ROS production and LKB1-AMPK activation, which reduces 26S proteasome-dependent degradation of p27(Kip1).