Inhibition of TRPM7 by carvacrol suppresses glioblastoma cell proliferation, migration and invasion.
Inhibition of TRPM7 by carvacrol suppresses glioblastoma cell proliferation, migration and invasion.
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DOI：
10.18632/oncotarget.3872
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发表时间：
2015-06-30
期刊：
影响因子：
--
通讯作者：
Sun HS
中科院分区：
文献类型：
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作者：
Chen WL;Barszczyk A;Turlova E;Deurloo M;Liu B;Yang BB;Rutka JT;Feng ZP;Sun HS
胶质母细胞瘤是具有破坏性增殖和侵袭特征的进行性脑肿瘤。离子通道是药物开发的第二大目标类别。在这项研究中，我们研究了TRPM 7抑制剂香芹酚对人U87胶质母细胞瘤细胞系的生存力，抗凋亡，迁移和侵袭力的影响。采用RT-PCR、Western blotting和免疫荧光法检测U87细胞中TRPM 7 mRNA和蛋白的表达水平。使用全细胞膜片钳技术记录TRPM 7电流。MTT测定用于评估细胞活力和增殖。伤口愈合和transwell实验用于评估细胞迁移和侵袭。检测p-Akt/t-Akt、p-ERK 1/2/t-ERK 1/2、caspase-3、MMP-2和磷酸化cofilin蛋白水平。TRPM 7 mRNA和蛋白在U87细胞中的表达高于正常人星形胶质细胞。全细胞膜片钳记录显示香芹酚阻断HEK 293细胞中的重组TRPM 7电流和U87细胞中的内源性TRPM 7样电流。香芹酚处理降低了U87细胞的活力、迁移和侵袭能力。香芹酚还降低MMP-2蛋白表达，促进cofilin的磷酸化。香芹酚还能抑制Ras/MEK/MAPK和PI 3 K/Akt信号通路。因此，香芹酚可能通过抑制TRPM 7通道而具有治疗胶质母细胞瘤的治疗潜力。
Glioblastomas are progressive brain tumors with devastating proliferative and invasive characteristics. Ion channels are the second largest target class for drug development. In this study, we investigated the effects of the TRPM7 inhibitor carvacrol on the viability, resistance to apoptosis, migration, and invasiveness of the human U87 glioblastoma cell line. The expression levels of TRPM7 mRNA and protein in U87 cells were detected by RT-PCR, western blotting and immunofluorescence. TRPM7 currents were recorded using whole-cell patch-clamp techniques. An MTT assay was used to assess cell viability and proliferation. Wound healing and transwell experiments were used to evaluate cell migration and invasion. Protein levels of p-Akt/t-Akt, p-ERK1/2/t-ERK1/2, cleaved caspase-3, MMP-2 and phosphorylated cofilin were also detected. TRPM7 mRNA and protein expression in U87 cells is higher than in normal human astrocytes. Whole-cell patch-clamp recording showed that carvacrol blocks recombinant TRPM7 current in HEK293 cells and endogenous TRPM7-like current in U87 cells. Carvacrol treatment reduced the viability, migration and invasion of U87 cells. Carvacrol also decreased MMP-2 protein expression and promoted the phosphorylation of cofilin. Furthermore, carvacrol inhibited the Ras/MEK/MAPK and PI3K/Akt signaling pathways. Therefore, carvacrol may have therapeutic potential for the treatment of glioblastomas through its inhibition of TRPM7 channels.