Li Yih-Do, Chou Chiang-Ting, Liang Wei-Zhe, Tseng Hui-Wen, Fang Yi-Chien, Hung Tzu-Yi, Chang Hong-Tai, Kuo Daih-Huang, Kuo Chun-Chi, Ho Chin-Man, Shieh Pochuen, Chen Fu-An, Jan Chung-Ren
Department of Laboratory Medicine, Zuoying Armed Forces General Hospital, Kaohsiung 81345, 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. 2015 Oct 31;58(5):285-93. doi: 10.4077/CJP.2015.BAD311.
The effect of the anti-inflammatory compound NPC-14686 on intracellular Ca²⁺ concentration (Ca²⁺) and viability in OC2 human oral cancer cells was investigated. The Ca²⁺-sensitive fluorescent probe fura-2 was used to examine Ca²⁺. NPC-14686 induced Ca²⁺ rises in a concentration-dependent fashion. The effect was reduced approximately by 10% by removing extracellular Ca²⁺. NPC-14686- elicited Ca²⁺ signal was decreased by nifedipine, econazole, SKF96365, and GF109203X. In Ca²⁺-free medium, incubation with the endoplasmic reticulum Ca²⁺ pump inhibitor thapsigargin or 2,5-di-tert-butylhydroquinone (BHQ) abolished NPC-14686-induced Ca²⁺ rises. Conversely, pretreatment with NPC-14686 abolished thapsigargin or BHQ-induced Ca²⁺ rises. Inhibition of phospholipase C with U73122 abolished NPC-14686-induced Ca²⁺ rises. At 20-100 μM, NPC-14686 inhibited cell viability, which was not reversed by chelating cytosolic Ca²⁺ with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'- tetraacetic acid-acetoxymethyl ester (BAPTA/AM). NPC-14686 between 20 μM and 40 μM also induced apoptosis. Collectively, in OC2 cells, NPC-14686 induced Ca²⁺ rises by evoking phospholipase C-dependent Ca²⁺ release from the endoplasmic reticulum and Ca²⁺ entry via protein kinase C-regulated store-operated Ca²⁺ channels. NPC-14686 also caused Ca²⁺-independent apoptosis.
研究了抗炎化合物NPC-14686对OC2人口腔癌细胞内Ca²⁺浓度（Ca²⁺）和细胞活力的影响。使用Ca²⁺敏感荧光探针fura-2检测Ca²⁺。NPC-14686以浓度依赖方式诱导Ca²⁺升高。去除细胞外Ca²⁺后，该效应降低约10%。硝苯地平、益康唑、SKF96365和GF109203X可降低NPC-14686引发的Ca²⁺信号。在无Ca²⁺培养基中，与内质网Ca²⁺泵抑制剂毒胡萝卜素或2,5-二叔丁基对苯二酚（BHQ）孵育可消除NPC-14686诱导的Ca²⁺升高。相反，用NPC-14686预处理可消除毒胡萝卜素或BHQ诱导的Ca²⁺升高。用U73122抑制磷脂酶C可消除NPC-14686诱导的Ca²⁺升高。在20-100μM时，NPC-14686抑制细胞活力，用1,2-双（2-氨基苯氧基）乙烷-N,N,N',N'-四乙酸-乙酰氧甲酯（BAPTA/AM）螯合胞质Ca²⁺不能逆转该抑制作用。20μM至40μM之间的NPC-14686也诱导细胞凋亡。总体而言，在OC2细胞中，NPC-14686通过引发内质网中磷脂酶C依赖性Ca²⁺释放和通过蛋白激酶C调节的储存-操作性Ca²⁺通道使Ca²⁺内流来诱导Ca²⁺升高。NPC-14686还引起不依赖Ca²⁺的细胞凋亡。
