Horng Chi-Ting, Chou Chiang-Ting, Sun Te-Kung, Liang Wei-Zhe, Kuo Chun-Chi, Wang Jue-Long, Shieh Pochuen, Jan Chung-Ren
Department of Ophthalmology, Kaohsiung Armed Force General Hospital, Kaohsiung 80284, Taiwan, Republic of China.
Department of Nursing, Division of Basic Medical Sciences, Chang Gung University of Science and Technology, Chia-Yi 61363, Taiwan, Republic of China.
Chin J Physiol. 2017 Oct 31;60(5):275-283. doi: 10.4077/CJP.2017.BAG506.
Carvacrol, a monoterpenic phenol compound, has been shown to possess various biological effects in different models. However, the effect of carvacrol on intracellular Ca²⁺ and its related physiology in human prostate cancer is unknown. This study explored the effect of carvacrol on cytosolic free Ca²⁺ levels ([Ca²⁺]i) and viability in PC3 human prostate cancer cells. Fura-2, a Ca²⁺- sensitive fluorescent dye, was used to assess [Ca²⁺]i. Cell viability was measured by the detecting reagent WST-1. Carvacrol at concentrations of 200-800 μM caused [Ca²⁺]i rises in a concentration-dependent manner. Removal of extracellular Ca²⁺ reduced carvacrol’s effect by approximately 60%. Carvacrol-induced Ca²⁺ entry was confirmed by Mn²⁺ entry-induced quench of fura-2 fluorescence, and was inhibited by approximately 30% by nifedipine, econazole, SKF96365, and the protein kinase C (PKC) inhibitor GF109203X. In Ca²⁺-free medium, treatment with the endoplasmic reticulum Ca²⁺ pump inhibitor thapsigargin (TG) abolished carvacrol-induced [Ca²⁺]i rises. Treatment with carvacrol also abolished TG-induced [Ca²⁺]i rises. Carvacrol-induced Ca²⁺ release from the endoplasmic reticulum was abolished by inhibition of phospholipase C (PLC). Carvacrol killed cells at concentrations of 200-600 μM in a concentration-dependent fashion. Chelating cytosolic Ca²⁺ with BAPTA/AM did not prevent carvacrol’s cytotoxicity. Together, in PC3 cells, carvacrol induced [Ca²⁺]i rises by inducing PLC-dependent Ca²⁺ release from the endoplasmic reticulum and Ca²⁺ entry via PKC-sensitive store-operated Ca²⁺ channels and other unknown channels. Carvacrol also induced Ca²⁺-dissociated cell death.
香芹酚是一种单萜酚类化合物，已被证明在不同模型中具有多种生物学效应。然而，香芹酚对人前列腺癌细胞内钙离子及其相关生理学的影响尚不清楚。本研究探讨了香芹酚对PC3人前列腺癌细胞胞质游离钙离子水平（[Ca²⁺]i）和细胞活力的影响。使用钙离子敏感荧光染料Fura-2评估[Ca²⁺]i。通过检测试剂WST-1测量细胞活力。浓度为200 - 800μM的香芹酚以浓度依赖性方式引起[Ca²⁺]i升高。去除细胞外钙离子可使香芹酚的作用降低约60%。通过锰离子进入诱导的Fura-2荧光淬灭证实了香芹酚诱导的钙离子内流，硝苯地平、酮康唑、SKF96365和蛋白激酶C（PKC）抑制剂GF109203X可使其抑制约30%。在无钙培养基中，用内质网钙离子泵抑制剂毒胡萝卜素（TG）处理可消除香芹酚诱导的[Ca²⁺]i升高。用香芹酚处理也可消除TG诱导的[Ca²⁺]i升高。抑制磷脂酶C（PLC）可消除香芹酚诱导的内质网钙离子释放。浓度为200 - 600μM的香芹酚以浓度依赖性方式杀死细胞。用BAPTA/AM螯合胞质钙离子并不能阻止香芹酚的细胞毒性。总之，在PC3细胞中，香芹酚通过诱导PLC依赖的内质网钙离子释放以及通过PKC敏感的储存-操纵性钙离子通道和其他未知通道引起钙离子内流，从而导致[Ca²⁺]i升高。香芹酚还诱导钙离子解离的细胞死亡。
