摘要
目的探讨卵泡抑素(follistation,FST)在下咽癌(hypopharyngeal carcinoma,HPC)中的生物学作用。方法收集2010年8月至2017年12月皖南医学院第一附属医院60例HPC患者,通过定量聚合酶链反应(qRT-PCR)和免疫组织化学检测FST表达水平。采用qRT-PCR检测其在FaDu以及永生鼻咽上皮细胞系NP-69细胞中的水平。构建特异性沉默FST mRNA的慢病毒载体(shFST)并转染FaDu细胞,观察细胞增殖,流式细胞术分析细胞周期、凋亡和迁移能力,在体接种检测细胞成瘤。此外,使用PathScan应激和凋亡信号蛋白芯片以及qRT-PCR分析FST敲减效应的机制。结果与正常组织和NP-69细胞相比,FST在HPC组织和FaDu细胞中的表达更高,且其高表达与肿瘤TNM分期呈正相关;shFST的抑制作用显著降低了FaDu细胞的增殖和迁移能力;FST沉默增加了FaDu细胞的凋亡比例,并使细胞周期停滞在G0/G1期和S期;FST沉默还抑制了体内肿瘤的生长。结论FST基因经ERK1/2和caspase信号,参与了HPC的发生发展进程,可能成为HPC的潜在治疗靶标。
Objective:To investigat the effect of follistation(FST)expression on hypopharyngeal carcinoma(HPC)tissues and cell lines.Methods:A total of 60 patients with HPC in the Yijishan Hospital of Wannan Medical College from August 2010 to December 2017 were recruited for this study.Levels of FST mRNA and protein were measured by quantitative polymerase chain reaction(PCR)and immunohistochemistry in HPC tissue samples and by qPCR in the HPC FaDu cells,as well as immortal nasopharyngeal epithelial cell line NP-69 cells.After silencing the FST expression in FaDu cells using lentivirus-mediated siRNA that was specific for FST mRNA,cell proliferation was determined by the Celigo assay.Tumor growth was monitored in nude mice and viability was detennined by MTT assay.The ratio of cell cycle arrest and apoptosis was carried out by flow cytometry.The colony formation ability was performed using Giemsa staining.In addition,wound healing assay were performed for analysis of cell motility.Furthermore,the putative mechanism of the effect of FST knockdown was analyzed using PathScan stress and apoptosis signaling array kit,and qPCR.Results:FST expression was significantly higher in human HPC tissue and FaDu cells when compared with normal tissue and NP-69 cells.A higher expression of FST in HPC samples was positively correlated with advanced tumors.Moreover,FST knockdown by shRNA significantly decreased cell growth,colony formation,and migration.Furthermore,FST silencing increased the cell apoptosis percentage,and arrested cell cycle in the G0/G1 and S phases in FaDu cells.FST silencing also suppressed tumor growth in vivo and might involve upregulating ERK1/2 and caspase signals.Conclusion:Our results indicate that the FST gene is associated with HPC progression,and may serve as a potential therapeutic target for the treatment of HPC.
作者
葛亮
刘少峰
Ge Liang;Liu Shaofeng(Department of Otorhinolaryngology Head and Neck Surgery,Yijishan Hospital of Wannan Medical College,Wuhu 241001,China)
出处
《山东第一医科大学(山东省医学科学院)学报》
2021年第7期481-487,共7页
Journal of Shandong First Medical University & Shandong Academy of Medical Sciences
基金
皖南医学院中青年科研基金(WK2019F11)o。
关键词
卵泡抑素
下咽癌
迁移
侵袭
增殖
细胞凋亡
follistatin
hypopharyngeal carcinoma
migration and invasion
proliferation
apoptosis