Antitumor activity of 7RH, a discoidin domain receptor 1 inhibitor, alone or in combination with dasatinib exhibits antitumor effects in nasopharyngeal carcinoma cells
Antitumor activity of 7RH, a discoidin domain receptor 1 inhibitor, alone or in combination with dasatinib exhibits antitumor effects in nasopharyngeal carcinoma cells
复制标题
DOI：
10.3892/ol.2016.5088
复制
发表时间：
2016-11-01
期刊：
影响因子：
2.9
通讯作者：
Guan, Zhong
中科院分区：
文献类型：
--
作者：
Lu, Qiu-Ping;Chen, Wen-Dan;Guan, Zhong
盘状结构域受体(DDRs)的失调与多种肿瘤的发生有关，包括头颈癌、鼻咽癌、乳腺癌、卵巢癌和食道癌。此外，抑制DDR1活性的药物被认为对鼻咽癌(NPC)的治疗有用。本研究的目的是评价DDR1sfk抑制(3-(2-(pyrazolo(1，5-a)pyrimidin-6-yl)-ethynyl)benzamide化合物7RH在体内外对鼻咽癌细胞的作用，以及与src家族激酶抑制剂达沙替尼合用时的作用。用四甲基偶氮唑盐比色法检测7RH单独或与达沙替尼合用对细胞活力的影响，用流式细胞仪检测细胞凋亡率。用免疫印迹法检测鼻咽癌细胞系CNE1、CNE2、HONE1和SUNE1中细胞周期相关基因的相对蛋白表达水平。用细胞黏附实验评估细胞迁移情况。在此基础上，建立了CNE2鼻咽癌裸鼠移植瘤模型，观察了7RH单独或与达沙替尼合用对鼻咽癌细胞生长的抑制作用。最后，通过将DDR1特异性小干扰RNA导入CNE2细胞，实现了DDR1蛋白表达的下调。7RH能有效抑制鼻咽癌细胞增殖，诱导细胞凋亡。此外，7RH还下调了Janus kinas1(JAK1)/信号转导和转录激活因子(STAT3)信号通路，而上调了Ras/Raf/丝裂原激活蛋白激酶(MEK)/细胞外信号调节激酶(ERK)和磷脂酰肌醇3-激酶(PI3K)/AKT信号通路的活性。此外，7RH处理的鼻咽癌细胞中磷酸化SRC的表达水平增加，提示SRC通过激活PI3K/AKT信号通路在鼻咽癌细胞对7RH的抵抗中发挥重要作用。本研究结果表明，抑制DDR1和SFK可能为鼻咽癌患者提供一种潜在的治疗策略。
Dysregulation of the discoidin domain receptors (DDRs) has been implicated in the development of numerous types of tumors, including head and neck cancer, and nasopharyngeal, breast, ovarian and esophageal carcinomas. Furthermore, agents that inhibit DDR1 activity are hypothesized to be useful for the treatment of nasopharyngeal carcinoma (NPC). The aim of the present study was to evaluate the effect of the DDR1 inhibitory (3-(2-(pyrazolo(1,5-a)pyrimidin-6-yl)-ethynyl)benzamide compound, 7RH, in NPC cells both in vitro and in vivo, and its effect when used in combination with dasatinib, a SRC family kinase (SFK) inhibitor. The effects of 7RH alone or in combination with dasatinib on cell viability were assessed using MTT assays and apoptosis was detected by flow cytometry. In addition, western blotting was performed to analyze the relative protein expression levels of cell cycle-associated genes in human NPC cell lines (CNE1, CNE2, HONE1 and SUNE1). Cell migration was also assessed using cell adhesion assays. Furthermore, tumor xenografts of CNE2 NPC cells were established in nude mice and the growth inhibitory effects of 7RH treatment alone or in combination with dasatinib were evaluated. Finally, knockdown of DDR1 protein expression was achieved by transfection of CNE2 cells with DDR1-specific small interfering RNA. Treatment with 7RH effectively suppressed the proliferation and induced the apoptosis of NPC cells. In addition, the Janus kinase 1 (JAK1)/signal transducer and activator of transcription (STAT3) signaling pathway was downregulated by 7RH, whereas the activities of the Ras/Raf/mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) and phosphoinositide 3-kinase (PI3K)/AKT signaling pathways were upregulated in response to 7RH treatment. Furthermore, the expression levels of phosphorylated SRC were increased in NPC cells treated with 7RH; thus indicating that SRC exhibits a vital function in the resistance of NPC cells to 7RH via activation of the PI3K/AKT signaling pathway. The results of the present study indicate that DDR1 and SFK inhibition may present a potential therapeutic strategy for patients with NPC.