Involvement of ROS-alpha v beta 3 integrin-FAK/Pyk2 in the inhibitory effect of melatonin on U251 glioma cell migration and invasion under hypoxia.
Involvement of ROS-alpha v beta 3 integrin-FAK/Pyk2 in the inhibitory effect of melatonin on U251 glioma cell migration and invasion under hypoxia.
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缺氧条件下ROS-αvβ3整合素-FAK/Pyk2参与褪黑素对U251胶质瘤细胞迁移和侵袭的抑制作用
DOI：
10.1186/s12967-015-0454-8
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发表时间：
2015-03-20
影响因子：
7.4
通讯作者：
Li ZQ
中科院分区：
文献类型：
--
作者：
Xu CS;Wang ZF;Huang XD;Dai LM;Cao CJ;Li ZQ
背景褪黑激素是一种抗氧化剂，具有抗胶质瘤侵袭的作用。然而，对于肿瘤发生发展的重要微环境--缺氧条件下褪黑素对胶质瘤细胞迁移和侵袭的影响却知之甚少。此外，粘着斑激酶（FAK）和富含脯氨酸的酪氨酸激酶2（Pyk 2）与细胞迁移和侵袭密切相关。因此，我们研究了这些激酶及其相关信号转导在褪黑素调节人U251胶质瘤细胞缺氧行为中的可能作用。方法采用创伤愈合和transwell实验检测U251胶质瘤细胞体外迁移和侵袭能力。用荧光探针6-羧基-2 ′，7′-二氯二氢荧光素二乙酸酯（DCFH-DA）测定细胞内活性氧（ROS）的产生。免疫荧光实验和蛋白质印迹分析检测蛋白质表达水平。小干扰RNA（siRNA）被用来沉默特定的基因expressions.ResultsThe药理学浓度（1 mM）的褪黑素显着抑制人U251胶质瘤细胞在缺氧条件下的迁移和侵袭。褪黑素的抑制作用伴随着FAK和Pyk 2磷酸化的减少以及α v β 3（αvβ3）整合素表达的减少。此外，通过siRNA抑制αvβ3整合素可降低FAK/Pyk 2的磷酸化水平，并显示出与褪黑素相似的抗肿瘤作用，提示αvβ3整合素- FAK/Pyk 2通路参与了褪黑素的抗迁移和抗侵袭作用。同时发现褪黑激素处理降低了缺氧条件下培养的U251胶质瘤细胞中的ROS水平。结论褪黑素通过ROS-α v β 3 integrin-FAK/Pyk 2信号通路抑制U251胶质瘤细胞在缺氧条件下的迁移和侵袭能力。这为褪黑激素可能是一种潜在的靶向胶质瘤缺氧微环境的治疗分子提供了证据。
BackgroundMelatonin, a well-known antioxidant, has been shown to possess anti-invasive properties for glioma. However, little is known about the effect of melatonin on glioma cell migration and invasion under hypoxia, which is a crucial microenvironment for tumor progress. In addition, focal adhesion kinase (FAK) and proline-rich tyrosine kinase 2 (Pyk2) are closely associated with cell migration and invasion. Therefore, we investigated the possible role of these kinases and its related signaling in the regulation of human U251 glioma cells behavior by melatonin under hypoxia.MethodsThe abilities of migration and invasion of U251 glioma cells were determined by wound healing and transwell assay in vitro. The intracellular production of reactive oxygen species (ROS) was measured by using the fluorescent probe 6-carboxy-2′, 7′-dichorodihydrofluorescein diacetate (DCFH-DA). Immunofluorescence experiments and western blotting analysis were used to detect the expression level of protein. Small interfering RNAs (siRNA) was used to silence specific gene expression.ResultsThe pharmacologic concentration (1 mM) of melatonin significantly inhibited the migration and invasion of human U251 glioma cells under hypoxia. The inhibitory effect of melatonin was accompanied with the reduced phosphorylation of FAK and Pyk2, and decreased expression of alpha v beta 3 (αvβ3) integrin. Additionally, inhibition of αvβ3 integrin by siRNA reduced the phosphorylation of FAK/Pyk2 and demonstrated the similar anti-tumor effects as melatonin, suggesting the involvement of αvβ3 integrin- FAK/Pyk2 pathway in the anti-migratory and anti-invasive effect of melatonin. It was also found that melatonin treatment decreased the ROS levels in U251 glioma cells cultured under hypoxia. ROS inhibitor apocynin not only inhibited αvβ3 integrin expression and the phosphorylation levels of FAK and Pyk2, but also suppressed the migratory and invasive capacity of U251 glioma cells under hypoxia.ConclusionsThese results suggest that melatonin exerts anti-migratory and anti-invasive effects on glioma cells in response to hypoxia via ROS-αvβ3 integrin-FAK/Pyk2 signaling pathways. This provides evidence that melatonin may be a potential therapeutic molecule targeting the hypoxic microenvironment of glioma.
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影响因子：
5.6
作者：
Lin TH;Tan TW;Tsai TH;Chen CC;Hsieh TF;Lee SS;Liu HH;Chen WC;Tang CH
通讯作者：
Tang CH
整联蛋白抑制剂抑制贝伐单抗诱导的神经胶质瘤侵袭。
影响因子：
5
作者：
Ishida, Joji;Onishi, Manabu;Kurozumi, Kazuhiko;Ichikawa, Tomotsugu;Fujii, Kentaro;Shimazu, Yosuke;Oka, Tetsuo;Date, Isao
通讯作者：
Date, Isao
影响因子：
11.2
作者：
Knowles LM;Gurski LA;Engel C;Gnarra JR;Maranchie JK;Pilch J
通讯作者：
Pilch J
影响因子：
10.3
作者：
Alvarez-Garcia, Virginia;Gonzalez, Alicia;Cos, Samuel
通讯作者：
Cos, Samuel
影响因子：
14
作者：
Lin, Li-Jung;Grimme, Jill M.;Sun, Jie;Lu, Shaoying;Gai, Lisa;Cropek, Donald M.;Wang, Yingxiao
通讯作者：
Wang, Yingxiao