The effect of magnolol on Ca2+ homeostasis and its related physiology in human oral cancer cells
The effect of magnolol on Ca2+ homeostasis and its related physiology in human oral cancer cells
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DOI：
10.1016/j.archoralbio.2018.02.006
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发表时间：
2018-05-01
影响因子：
3
通讯作者：
Jan, Chung-Ren
中科院分区：
文献类型：
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作者：
Hsieh, Shu-Feng;Chou, Chiang-Ting;Jan, Chung-Ren
目的 厚朴酚是一种来自草药的多酚化合物，已被证明可以改变各种细胞模型的生理机能。然而，厚朴酚对口腔癌细胞中 Ca2+ 稳态及其相关生理学的影响尚不清楚。本研究检查厚朴酚是否改变 OC2 人口腔癌细胞中的 Ca2+ 信号传导和细胞活力。方法：使用荧光 Ca2+ 敏感染料 fura-2 测量悬浮细胞中的胞质 Ca2+ 浓度 ([Ca2+](i))。通过 4-[3-[4-lodopheny1]-2-4(4-硝基苯基1)-2H-5-tetrazolio-1,3-苯二磺酸]水溶性四唑-1 (WST-1) 测定法检查细胞活力。结果：浓度为 20-100 mu M 的厚朴酚诱导 [Ca2+](i) 升高。 Ca2+ 去除使信号减少约 50%。厚朴酚 (100 μM) 诱导 Mn2+ 流入，表明 Ca2+ 进入。厚朴酚诱导的 Ca2+ 内流受到蛋白激酶 C (PKC) 调节剂和钙池操纵的 C-a2+ 通道抑制剂的部分抑制。在无 Ca2+ 培养基中，用内质网 Ca2+ 泵抑制剂 2,5-二叔丁基氢醌 (BHQ) 处理可消除厚朴酚诱发的 [Ca2+](i) 升高。相反，用厚朴酚治疗消除了 BHQ 诱发的 [Ca2+](i) 上升。 U73122 抑制磷脂酶 C (PLC) 部分抑制厚朴酚诱导的 [Ca2+](i) 上升。 20-100 mu M 的厚朴酚会降低细胞活力，这种情况不能通过 Ca2+ 螯合剂 1,2-双（2-氨基苯氧基）乙烷-N,N,N',N'-四乙酸-乙酰氧基甲酯 (BAPTA/AM) 预处理来逆转。结论：在 OC2 细胞中，厚朴酚通过部分激发而诱导 [Ca2+](i) 升高。 PLC 依赖性 Ca2+ 从内质网释放，Ca2+ 通过 PKC 敏感的存储操纵的 Ca-2+ 进入。厚朴酚还引起不依赖 Ca2+ 的细胞死亡。因此，厚朴酚诱导的细胞毒性可能不参与口腔癌细胞中与细胞内 Ca2+ 动员相关的激活机制。
Objective Magnolol, a polyphenol compound from herbal medicines, was shown to alter physiology in various cell models. However, the effect of magnolol on Ca2+ homeostasis and its related physiology in oral cancer cells is unclear. This study examined whether magnolol altered Ca2+ signaling and cell viability in OC2 human oral cancer cells.Methods: Cytosolic Ca2+ concentrations ([Ca2+](i)) in suspended cells were measured by using the fluorescent Ca2+ -sensitive dye fura-2. Cell viability was examined by 4-[3-[4-lodopheny1]-2-4(4-nitropheny1)-2H-5-tetrazolio-1,3-benzene disulfonate] water soluble tetrazolium-1 (WST-1) assay.Results: Magnolol at concentrations of 20-100 mu M induced [Ca2+](i) rises. Ca2+ removal reduced the signal by approximately 50%. Magnolol (100 mu M) induced Mn2+ influx suggesting of Ca2+ entry. Magnolol-induced Ca2+ entry was partially suppressed by protein kinase C (PKC) regulators, and inhibitors of store-operated C-a2+ channels. In Ca2+-free medium, treatment with the endoplasmic reticulum Ca2+ pump inhibitor 2,5-di-tertbutylhydroquinone (BHQ) abolished magnolol-evoked [Ca2+](i) rises. Conversely, treatment with magnolol abolished BHQ-evoked [Ca2+](i) rises. Inhibition of phospholipase C (PLC) with U73122 partially inhibited magnolol-induced [Ca2+](i) rises. Magnolol at 20-100 mu M decreased cell viability, which was not reversed by pretreatment with the Ca2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester (BAPTA/AM).Conclusions: Together, in OC2 cells, magnolol induced [Ca2+](i) rises by evoking partially PLC-dependent Ca2+ release from the endoplasmic reticulum and Ca2+ entry via PKC-sensitive store-operated Ca-2+ entry. Magnolol also caused Ca2+-independent cell death. Therefore, magnolol-induced cytotoxicity may not be involved in activation mechanisms associated with intracellular Ca2+ mobilization in oral cancer cells.