Wang Jue-Long, Jan Chung-Ren, Chen Min-Huey
Department of Rehabilitation and Traditional Medicine, Kaohsiung Veterans General Hospital, Kaohsiung, Taiwan.
Department of Medical Education and Research, Kaohsiung Veterans General Hospital, Kaohsiung, Taiwan.
J Dent Sci. 2023 Jul;18(3):1280-1287. doi: 10.1016/j.jds.2023.02.013. Epub 2023 Mar 6.
BACKGROUND/PURPOSE: Econazole is an antifungal drug. Antifungal activity of econazole against non-dermatophyte molds was reported. Econazole inhibited Ca channels and stimulated cytotoxicity in lymphoma and leukemia cells. Ca cations are crucial second envoy that triggers various processes. This research was aimed to investigate action of econazole on Ca levels and cytotoxicity in OC2 human oral cancer cells.
Cytosolic Ca levels ([Ca]) were detected employing fura-2 as a probe in a RF-5301PC spectrofluorophotometer (Shimadzu). Cytotoxicity was determined using 4-[3-[4-lodophenyl]-2-4(4-nitrophenyl)-2H-5-tetrazolio-1,3-benzene disulfonate] (WST-1) to detect fluorescence changes.
Econazole at 10-50 μmol/L provoked [Ca] raises. Forty % of 50 μml//L econazole-induced signal was diminished when external Ca was eliminated. The Ca influx provoked by econazole was suppressed by different degrees by store-induced Ca influx suppressors SKF96365 and nifedipine; GF109203X (a protein C [PKC] inhibitor); an extracellular signaling pathway (ERK) 1/2 blocker PD98059, and phospholipase A2 suppressor aristolochic acid, but was enhanced by phorbol 12-myristate 13 acetate (PMA; a PKC activator) by 18%. Without external Ca, econazole-caused [Ca] raises were abolished by thapsigargin. In contrast, econazole partially suppressed the [Ca] raises caused by thapsigargin. U73122 fell short to change econazole-caused [Ca] responses. Econazole (10-70 μmol/L) elicited cytotoxicity in a dose-dependent fashion. Blockade of 50 μmol/L econazole-induced [Ca] rises with BAPTA/AM enhanced econazole-induced cytotoxicity by 72%.
Econazole evoked [Ca] raises and provoked cytotoxicity in a concentration-dependent manner in OC2 human oral cancer cells. In Ca-containing solution, BAPTA/AM enhanced 50 μmol/L econozole-induced cytotoxicity.
背景/目的：益康唑是一种抗真菌药物。有报道称益康唑对非皮肤癣菌霉菌具有抗真菌活性。益康唑可抑制钙通道并刺激淋巴瘤和白血病细胞的细胞毒性。钙离子是触发各种过程的关键第二信使。本研究旨在探讨益康唑对OC2人口腔癌细胞中钙水平和细胞毒性的作用。
使用fura-2作为探针，在RF-5301PC分光荧光光度计（岛津）中检测胞质钙水平（[Ca]）。使用4-[3-[4-碘苯基]-2-4(4-硝基苯基)-2H-5-四氮唑]-1,3-苯二磺酸盐（WST-1）检测荧光变化来确定细胞毒性。
10 - 50 μmol/L的益康唑可引起[Ca]升高。当去除细胞外钙时，50 μmol/L益康唑诱导的信号减弱了40%。益康唑引发的钙内流在不同程度上受到储存诱导的钙内流抑制剂SKF96365和硝苯地平、蛋白C（PKC）抑制剂GF109203X、细胞外信号调节激酶（ERK）1/2阻滞剂PD98059以及磷脂酶A2抑制剂马兜铃酸的抑制，但佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA；一种PKC激活剂）可使其增强18%。在没有细胞外钙的情况下，毒胡萝卜素可消除益康唑引起的[Ca]升高。相反，益康唑可部分抑制毒胡萝卜素引起的[Ca]升高。U73122未能改变益康唑引起的[Ca]反应。益康唑（10 - 70 μmol/L）以剂量依赖性方式引起细胞毒性。用BAPTA/AM阻断50 μmol/L益康唑诱导的[Ca]升高可使益康唑诱导的细胞毒性增强72%。
益康唑在OC2人口腔癌细胞中以浓度依赖性方式引起[Ca]升高并引发细胞毒性。在含钙溶液中，BAPTA/AM增强了50 μmol/L益康唑诱导的细胞毒性。
