长链非编码RNAALOX12P2对口腔鳞状细胞癌细胞活力、迁移及侵袭的影响  

Effect of long noncoding RNA ALOX12P2 on viability,migration and invasion of oral squamous cell carcinoma cells

作　　者：公慧 朱靖 郑文甜 郝嘉仪 王亚杰 蒋英英 GONG Hui;ZHU Jing;ZHENG Wentian;HAO Jiayi;WANG Yajie;JIANG Yingying(School of Stomatology,Shandong Second Medical University,Weifang 261053,China;Department of Dentistry,Affiliated Hospital of Shandong Second Medical University,Weifang 261035,China)

机构地区：[1]山东第二医科大学口腔医学院,山东潍坊261053 [2]山东第二医科大学附属医院口腔科,山东潍坊261053

出　　处：《中国病理生理杂志》2024年第11期2031-2040,共10页Chinese Journal of Pathophysiology

基　　金：山东省自然科学基金资助项目(No.ZR2023LSW019);2021年山东省高等学校“青创人才引育计划”支持项目(No.2021-5);山东省大学生创新创业训练计划项目(No.S202410438019);潍坊医学院研究生科研创新基金资助项目(No.2023YJSCX005);山东第二医科大学校级大学生创新创业训练计划项目(No.X2024081,No.X2024082)

摘　　要：目的:本研究旨在深入了解ALOX12P2在口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)中的表达和定位,以及其对口腔鳞状细胞癌细胞活力、迁移及侵袭产生的影响。方法:(1)通过数据库UALCAN(the University of Alabama at Birmingham cancer data analysis portal),对头颈部鳞状细胞癌(head and neck squamous cell carcinoma,HNSCC)组织中ALOX12P2的表达及其与临床病理特征的相关性进行分析;利用GDC和UCSC Xena数据库分析ALOX12P2在OSCC中的表达和对生存预后的影响。(2)对OSCC细胞系中ALOX12P2的表达进行实时荧光定量PCR(RT-qPCR)检测。(3)ALOX12P2的亚细胞定位通过RNA核质分离实验检测。(4)对CAL-27细胞建立AL-OX12P2敲减组(SS-ALOX12P2组)和敲减对照组(SS-NC组);对HN30细胞建立ALOX12P2过表达组(ALOX12P2组)和过表达对照组(vector组);由CCK-8和Transwell评估ALOX12P2对细胞活力、迁移和侵袭能力的影响。(5)由Western blot实验检测ALOX12P2表达水平改变后对上皮-间充质转化(epithelial-mesenchymal transition,EMT)相关基因以及PI3K/AKT通路的影响。结果:与正常组织相比,ALOX12P2在HNSCC和OSCC组织中的表达较高,其表达与不良预后有关;RT-qPCR结果显示,在6种OSCC细胞系中,ALOX12P2的相对表达水平均高于正常细胞(P<0.05);RNA核质分离结果显示,ALOX12P2定位于细胞核;与SS-NC组比较,SS-ALOX12P2组中ALOX12P2相对表达量明显降低(P<0.01),细胞活力、迁移及侵袭能力也明显降低(P<0.01);与vector组相比,ALOX12P2组中AL-OX12P2相对表达量明显增高(P<0.01),细胞活力、迁移及侵袭能力也明显升高(P<0.01);Western blot结果显示,敲减ALOX12P2导致E-钙粘蛋白(E-cadherin)的蛋白水平升高,N-钙粘蛋白(N-cadherin)和波形蛋白(Vimentin)的蛋白水平降低(P<0.01);ALOX12P2过表达而结果显示相反(P<0.01);敲减ALOX12P2导致p-PI3K和p-AKT的蛋白表达减少(P<0.01),过表达ALOX12P2则显示p-PI3K和p-AKT的蛋白表达增多(P<0.01)。结论:在OSCC中,ALOX12P2�AIM:This study aimed to investigate the expression and localization of ALOX12P2 in oral squa-mous cell carcinoma(OSCC),as well as its effects on cell viability,migration,and invasion.METHODS:The expres-sion of ALOX12P2 in head and neck squamous cell carcinoma(HNSCC)tissues and its correlation with clinicopathologi-cal features were analyzed using the UALCAN database(University of Alabama at Birmingham Cancer Data Analysis Por-tal).Additionally,the expression of ALOX12P2 in OSCC and its impact on survival prognosis were evaluated through the GDC and UCSC Xena databases.The expression levels of ALOX12P2 in OSCC cell lines were assessed via quantitative re-al-time PCR(RT-qPCR).The subcellular localization of ALOX12P2 was determined using nucleoplasmic RNA isola-tion.CAL-27 cells were used to establish an ALOX12P2 knockdown group(SS-ALOX12P2)and a control group(SS-NC).HN30 cells were employed to form an ALOX12P2 overexpression group(ALOX12P2)and a control group(vector).The effects of altered ALOX12P2 expression on the epithelial-mesenchymal transition(EMT)-related gene E-cadherin and the PI3K/AKT signaling pathway were assessed through Western blot analysis.RESULTS:ALOX12P2 expression was significantly higher in HNSCC and OSCC tissues compared to normal tissues,with its expression correlating with poor prog-nosis.RT-qPCR analysis indicated that the relative expression of ALOX12P2 in OSCC cells was comparable to that in nor-mal cells(P<0.05).RNA nucleoplasmic isolation confirmed that ALOX12P2 localized in the nucleus.In comparison to the SS-NC group,the SS-ALOX12P2 group exhibited a marked reduction in ALOX12P2 expression(P<0.01),alongside significant decreases in cell viability,migration,and invasion(P<0.01).Conversely,the ALOX12P2 group showed sub-stantially higher relative expression compared to the vector group(P<0.01),with enhanced cell viability,migration,and invasion abilities(P<0.01).Western blot analysis demonstrated that ALOX12P2 knockdown resulted in upregulation of E-cadherin and downregulation of N-cadher

关 键 词：口腔鳞状细胞癌 长链非编码RNA 细胞活力 细胞迁移 细胞侵袭 

分 类 号：R739.8[医药卫生—肿瘤] Q279[医药卫生—临床医学] R363[生物学—细胞生物学]