Su Ching-Kai, Chou Chiang-Ting, Lin Ko-Long, Liang Wei-Zhe, Cheng Jin-Shiung, Chang Hong-Tai, Chen I-Shu, Lu Ti, Kuo Chun-Chi, Yu Chia-Cheng, Shieh Pochuen, Kuo Daih-Huang, Chen Fu-An, Jan Chung-Ren
a Department of Medicine , Kaohsiung Veterans General Hospital Pingtung Branch , Pingtung , Taiwan.
b Department of Nursing, Division of Basic Medical Sciences , Chang Gung Institute of Technology , Chia-Yi , Taiwan.
Toxicol Mech Methods. 2016 Oct;26(8):580-587. doi: 10.1080/15376516.2016.1216208. Epub 2016 Oct 28.
Tricyclic antidepressants (TCA) have been clinically prescribed in the auxiliary treatment of cancer patients. Although protriptyline, a type of TCA, was used primarily in the clinical treatment of mood disorders in cancer patients, the effect of protriptyline on physiology in human osteosarcoma is unknown. This study examined the effect of protriptyline on cytosolic free Caconcentrations ([Ca]) and viability in MG63 human osteosarcoma cells. Protriptyline between 50 and 250 μM evoked [Ca] rises concentration-dependently. Protriptyline induced influx of Mn, indirectly implicating Cainflux. Protriptyline-evoked Caentry was inhibited by nifedipine by 20% but was not altered by econazole, SKF96365, GF109203X, and phorbol-12-myristate-13-acetate (PMA). In Ca-free medium, treatment with protriptyline inhibited the endoplasmic reticulum Capump inhibitor thapsigargin-evoked [Ca] rises. Conversely, treatment with thapsigargin inhibited 45% of protriptyline-evoked [Ca] rises. Inhibition of phospholipase C (PLC) with U73122 failed to alter protriptyline-evoked [Ca] rises. Protriptyline at 50-250 μM decreased cell viability, which was not reversed by pretreatment with the Cachelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester (BAPTA/AM). Collectively, our data suggest that in MG63 cells, protriptyline induced [Ca] rises by evoking Carelease from the endoplasmic reticulum and other stores in a PLC-independent manner, and Caentry via a nifedipine-sensitive Capathway. Protriptyline also caused Ca-independent cell death.
三环类抗抑郁药（TCA）已被临床用于癌症患者的辅助治疗。尽管作为TCA一种的普罗替林主要用于癌症患者情绪障碍的临床治疗，但其对人骨肉瘤生理学的影响尚不清楚。本研究检测了普罗替林对MG63人骨肉瘤细胞胞质游离钙浓度（[Ca]）和活力的影响。50至250μM的普罗替林浓度依赖性地引起[Ca]升高。普罗替林诱导锰内流，间接提示钙内流。硝苯地平可抑制普罗替林诱发的钙内流20%，但酮康唑、SKF96365、GF109203X和佛波醇-12-肉豆蔻酸酯-13-乙酸酯（PMA）对其无影响。在无钙培养基中，普罗替林处理可抑制内质网钙泵抑制剂毒胡萝卜素诱发的[Ca]升高。相反，毒胡萝卜素处理可抑制45%的普罗替林诱发的[Ca]升高。用U73122抑制磷脂酶C（PLC）未能改变普罗替林诱发的[Ca]升高。50至250μM的普罗替林可降低细胞活力，用钙螯合剂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）预处理不能逆转这一作用。总体而言，我们的数据表明，在MG63细胞中，普罗替林通过以PLC非依赖方式诱发内质网和其他储存库释放钙以及通过硝苯地平敏感的钙通道引起钙内流，从而诱导[Ca]升高。普罗替林还导致非钙依赖性细胞死亡。
