Zhuang Ya-Jing, Liao Zhi-Wei, Yu Hong-Wei, Song Xian-Lu, Liu Yuan, Shi Xing-Yuan, Lin Xiao-Dan, Zhou Tong-Chong
a Department of Radiation Oncology ; Cancer Center of Guangzhou Medical University ; Guangzhou , China.
Cancer Biol Ther. 2015;16(1):88-96. doi: 10.4161/15384047.2014.987029.
Ubiquitin-specific protease 22 (USP22) is closely related with poor prognosis of cancer patients. However, the role of USP22 expression in nasopharyngeal carcinoma (NPC) has not been determined. The main aim of this study was to determine the role of USP22 in the pathologic processes of NPC. Immunohistochemistry (IHC), western blot (WB), and real-time polymerase chain reaction (RT-PCR) were used to measure the expression of USP22 in cell lines and tissues of NPC in comparison with expression in non-cancerous cells and tissues. USP22-specific short hairpin RNA (shRNA) was used to knock down USP22 expression in the NPC cell line CNE-1 and CNE-2. Furthermore, the impact of USP22 in cellular proliferation, growth, and cell cycle were detected respectively. WB was used to determine the role of USP22 in the AKT/GSK-3/Cyclin signaling pathway. The expression levels of USP22 were remarkably higher in NPC cell lines and tissues. With cell counting and the MTS assay, cellular growth and proliferation progression of USP22 knockdown cell line was shown to be effectively restrained. The USP22 silencing both in CNE-1 and CNE-2 cells caused them to accumulate in the G0/G1 phase of the cell cycle. USP22 knockdown was also found to modulate the AKT/GSK-3/Cyclin pathway, resulting in downregulation of p-AKT, p-GSK-3β, and cyclinD1. This study suggests that USP22 plays a critical regulatory role in the pathologic processes of NPC, and that it may be a potential biological treatment target in the future.
泛素特异性蛋白酶22(USP22)与癌症患者的不良预后密切相关。然而,USP22表达在鼻咽癌(NPC)中的作用尚未确定。本研究的主要目的是确定USP22在NPC病理过程中的作用。与非癌细胞和组织中的表达相比,采用免疫组织化学(IHC)、蛋白质免疫印迹(WB)和实时聚合酶链反应(RT-PCR)来检测NPC细胞系和组织中USP22 的表达。使用USP22特异性短发夹RNA(shRNA)敲低NPC细胞系CNE-1和CNE-2中USP22的表达。此外,分别检测了USP22对细胞增殖、生长和细胞周期的影响。采用WB确定USP22在AKT/GSK-3/细胞周期蛋白信号通路中的作用。USP22在NPC细胞系和组织中的表达水平显著更高。通过细胞计数和MTS试验,结果显示USP22敲低细胞系的细胞生长和增殖进程受到有效抑制。在CNE-1和CNE-2细胞中,USP22沉默均导致它们在细胞周期的G0/G1期积累。还发现USP22敲低可调节AKT/GSK-3/细胞周期蛋白通路,导致p-AKT、p-GSK-3β和细胞周期蛋白D1下调。本研究表明,USP22在NPC的病理过程中起关键调节作用,并且它可能是未来潜在的生物治疗靶点。