机构地区:[1]潍坊医学院口腔医学院,山东潍坊261053 [2]潍坊医学院附属医院口腔科,山东潍坊261035
出 处:《中国癌症杂志》2023年第1期45-53,共9页China Oncology
基 金:山东省自然科学基金(ZR2020MH192);潍坊医学院博士启动基金(04103801);2021年山东省高等学校“青创人才引育计划”(基于多组学口腔癌分子流行病学研究创新团队)。
摘 要:背景与目的:口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)是头颈部鳞状细胞癌中最普遍的亚型,其发病机制尚不清楚。核仁蛋白8(nucleolar protein 8,NOL8)作为RNA结合蛋白(RNA-binding protein,RBP),在多种肿瘤的发生、发展中发挥关键作用,但其在OSCC中的作用尚不清楚,本研究旨在探讨NOL8在OSCC中的表达水平,并研究其对OSCC细胞增殖、迁移、侵袭和上皮-间充质转化(epithelial-mesenchymal transition,EMT)的影响。方法:利用基因表达谱交互分析2(gene expression profiling interactive analysis 2,GEPIA2)、肿瘤免疫评估资源(tumor immune estimation resource,TIMER)、阿拉巴马大学伯明翰分校癌症数据分析(the University of Alabama at Birmingham cancer data analysis portal,UALCAN)及RNA相互作用数据库(the encyclopedia of RNA interactomes,ENCORI)在线分析NOL8在头颈鳞癌组织中的表达;采用实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR)检测NOL8在OSCC细胞中的mRNA表达水平。利用siRNA干扰技术下调NOL8在CAL-27细胞中的表达,形成NOL8敲低组(siNOL8-1,si-NOL8-2)及阴性对照组;通过慢病毒转染技术感染CAL-27及HN6细胞过表达NOL8,形成NOL8过表达组及阴性对照组。分别通过细胞计数试剂盒-8(cell counting kit-8,CCK-8)实验、划痕愈合试验及transwell实验检测NOL8表达水平变化后对OSCC细胞增殖、迁移和侵袭的影响;采用蛋白质印迹法(Western blot)检测NOL8表达水平改变后对EMT相关基因E-钙黏蛋白(E-cadherin)、波形蛋白(vimentin)和N-钙黏蛋白(N-cadherin)表达的影响。利用裸鼠皮下移植瘤模型探究NOL8在体内对OSCC细胞增殖的影响。结果:GEPIA2、TIMER、UALCAN及ENCORI在线分析显示,与头颈鳞癌的癌旁正常组织相比,NOL8在头颈鳞癌组织中表达升高;RTFQ-PCR结果显示,与正常对照细胞相比,NOL8在OSCC细胞系中的mRNA表达显著上调。转染si-NOL8-1及si-NOL8-2的CAL-27细�Background and purpose:Oral squamous cell carcinoma(OSCC)is the most common subtype of head and neck squamous cell carcinoma(HNSCC),and its pathogenesis is unclear.Nucleolar protein 8(NOL8),as one of the RNA-binding protein(RBP),plays a key role in the occurrence and development of many kinds of tumors,however its role in OSCC is not clear.This study aimed to investigate the expression level of NOL8 in OSCC and its effects on the proliferation,migration,invasion and epithelial-mesenchymal transition(EMT)of OSCC.Methods:The expression of NOL8 in HNSCC was analyzed online by gene expression profiling interactive analysis 2(GEPIA2),tumor immune estimation resource(TIMER),the University of Alabama at Birmingham cancer data analysis portal(UALCAN)and the encyclopedia of RNA interactomes(ENCORI).The mRNA expression level of NOL8 in OSCC cells was detected by real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR).siRNA interference technique was used to knock down the expression of NOL8 in CAL-27 cells to form NOL8 knockdown group(siNOL8-1,si-NOL8-2)and negative control group.CAL-27 and HN6 cells were overexpressed with NOL8 by lentivirus transfection technique to form NOL8 overexpression group and negative control group.Cell counting kit-8(CCK-8)assay,scratch healing assay and transwell assay were used to detect the effect of NOL8 expression on the proliferation,migration and invasion of OSCC cells.Western blot assay was used to detect the effect of NOL8 on the expression of EMT-related genes including E-cadherin,vimentin and N-cadherin.The effect of NOL8 on the proliferation of OSCC cells in vivo was examed by xenograft formation assays.Results:The online analysis of GEPIA2,TIMER,UALCAN and ENCORI showed that the expression of NOL8 was higher in HNSCC than in normal tissues,and the expression of NOL8 in OSCC was significantly higher than in normal control cells.The relative expression of NOL8 in CAL-27 cells transfected with si-NOL8-1 and si-NOL8-2 was significantly lower compared with the negative co
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