Wang Limei, Wu Xiuyin, Wang Ruolin, Yang Chengzhe, Li Zhi, Wang Cunwei, Zhang Fenghe, Yang Pishan
Department of Periodontology, School of Stomatology, Shandong University, 44-1 West Wen Hua Road, Jinan, 250012, Shandong, People's Republic of China.
Shandong Provincial Key Laboratory of Oral Tissue Regeneration, Jinan, 250012, Shandong, China.
Biol Res. 2017 May 25;50(1):19. doi: 10.1186/s40659-017-0124-9.
Bromodomain-containing protein 4 (BRD4) inhibition is a new therapeutic strategy for many malignancies. In this study, we aimed to explore the effect of BRD4 inhibition by JQ1 on in vitro cell growth, migration and invasion of salivary adenoid cystic carcinoma (SACC).
The human normal epithelial cells and SACC cells (ACC-LM and ACC-83) were treated with JQ1 at concentrations of 0, 0.1, 0.5 or 1 μM. Cell Counting Kit-8 (CCK-8) assay was performed to evaluate cell proliferation. Cell apoptosis and cell cycle distribution was evaluated by Flow cytometry. Immunofluorescence staining was used to examine the expression of BRD4 in SACC cells. The quantitative real-time polymerase chain reaction (qRT-PCR) assay and western blot assay were performed to examine messenger RNA (mRNA) and protein levels in SACC cells. Wound-healing assay and transwell assay were used to evaluate the activities of migration and invasion of SACC cells.
JQ1 exhibits no adverse effects on proliferation, cell cycle and cell apoptosis of the normal human epithelial cells, while suppressed proliferation and cell cycle, and induced apoptosis of SACC cells, down-regulated the mRNA and protein levels of BRD4 in SACC cells, meanwhile reduced protein expressions of c-myc and BCL-2, two known target genes of BRD4. Moreover, JQ1 inhibited SACC cell migration and invasion by regulating key epithelial-mesenchymal transition (EMT) characteristics including E-cadherin, Vimentin and Twist.
BRD4 is an important transcription factor in SACC and BRD4 inhibition by JQ1 may be a new strategy for SACC treatment.
含溴结构域蛋白4（BRD4）抑制是多种恶性肿瘤的一种新治疗策略。在本研究中，我们旨在探讨JQ1抑制BRD4对涎腺腺样囊性癌（SACC）体外细胞生长、迁移和侵袭的影响。
将人正常上皮细胞和SACC细胞（ACC-LM和ACC-83）用浓度为0、0.1、0.5或1 μM的JQ1处理。采用细胞计数试剂盒-8（CCK-8）法评估细胞增殖。通过流式细胞术评估细胞凋亡和细胞周期分布。免疫荧光染色用于检测SACC细胞中BRD4的表达。采用定量实时聚合酶链反应（qRT-PCR）法和蛋白质免疫印迹法检测SACC细胞中的信使核糖核酸（mRNA）和蛋白质水平。采用伤口愈合试验和Transwell试验评估SACC细胞的迁移和侵袭活性。
JQ1对正常人上皮细胞的增殖、细胞周期和细胞凋亡无不良影响，而抑制了SACC细胞的增殖和细胞周期，并诱导其凋亡，下调了SACC细胞中BRD4的mRNA和蛋白质水平，同时降低了BRD4的两个已知靶基因c-myc和BCL-2的蛋白质表达。此外，JQ1通过调节关键的上皮-间质转化（EMT）特征，包括E-钙黏蛋白、波形蛋白和Twist，抑制SACC细胞的迁移和侵袭。
BRD4是SACC中的一种重要转录因子，JQ1抑制BRD4可能是SACC治疗的一种新策略。
