miR-4465对喉癌Hep-2细胞增殖、侵袭和迁移的影响及其机制  

Effect of miR-4465 on proliferation,invasion and migration of laryngeal cancer Hep-2 cells and its mechanism

作　　者：高天乐 郭志强 孙青 刘建 李俊 GAO Tianle;GUO Zhiqiang;SUN Qing;LIU Jian;LI Jun(Department of Otolaryngology,Qingpu Branch of Zhongshan Hospital Affi-liated to Fudan University,Shanghai 201700,China)

机构地区：[1]复旦大学附属中山医院青浦分院耳鼻喉科,上海201700

出　　处：《山西医科大学学报》2023年第10期1347-1353,共7页Journal of Shanxi Medical University

基　　金：上海市青浦区卫生健康委员会基金项目(W2018-01)。

摘　　要：目的探讨miR-4465调控第10号染色体同源丢失性磷酸酶-张力蛋白基因(phosphatase and tensin homolog deleted on chromosome 10,PTEN)甲基化对喉癌Hep-2细胞增殖、侵袭和迁移的影响。方法选取2017年1月至2022年1月复旦大学附属中山医院青浦分院耳鼻喉科手术治疗患者的喉癌组织及相应的癌旁组织20例,实时荧光定量RT-PCR检测组织中miR-4465和PTEN的表达,甲基化特异性PCR检测PTEN甲基化水平,Pearson法分析癌组织中miR-4465和PTEN的相关性,并分析PTEN基因启动子甲基化率与性别、年龄、临床分型、肿瘤分期、淋巴结转移的关系。将Hep-2细胞分为空白组、对照组(转染mimic NC)、miR-4465 mimic组(转染miR-4465 mimic),qRT-PCR检测Hep-2细胞中PTEN mRNA表达,Western blot检测Hep-2细胞中PTEN蛋白表达,CCK-8检测Hep-2细胞增殖,Transwell实验检测Hep-2细胞侵袭,细胞划痕愈合实验检测Hep-2细胞迁移。再将Hep-2细胞分为pcDNA3.0组(转染pcDNA3.0质粒)和pcDNA3.0-PTEN组(转染pcDNA3.0-PTEN质粒),CCK-8检测细胞增殖,Transwell实验检测细胞侵袭,细胞划痕愈合实验检测细胞迁移。结果喉癌组织中miR-4465和PTEN的相对表达低于癌旁组织(P<0.001);癌组织中miR-4465的表达与PTEN呈正相关性(r=0.459,P=0.036);PTEN在癌组织的甲基化率明显高于癌旁组织(P<0.001);PTEN基因启动子甲基化率与性别、年龄、临床分型无相关性(P>0.05),与肿瘤分期、淋巴结转移具有一定的关系(P<0.001)。与空白组和对照组相比,miR-4465 mimic组PTEN mRNA和蛋白相对表达显著升高(P<0.001),miR-4465 mimic组Hep-2细胞增殖能力明显降低(P<0.001),Hep-2细胞侵袭能力明显降低(P<0.001),Hep-2细胞迁移率明显降低(P<0.001)。与pcDNA3.0组相比,pcDNA3.0-PTEN组Hep-2细胞增殖、侵袭、迁移能力明显降低(P<0.001)。结论miR-4465在喉癌组织中低表达,过表达miR-4465可以抑制Hep-2细胞的增殖、侵袭和迁移,这可能与PTEN启动子甲基化程度降低和PTENObjective To investigate the effect of miR-4465 regulating homologous loss phosphatase tensin gene(PTEN)methylation on the proliferation,invasion,and migration abilities of laryngeal cancer Hep-2 cells.Methods Twenty cases of laryngeal cancer tissues and their corresponding paracancer tissues from patients from January 2017 to January 2022 in Department of Otolaryngology,Qingpu Branch of Zhongshan Hospital Affiliated to Fudan University were collected.Real time fluorescence quantitative RT-PCR was used to detect the expression of miR-4465 and PTEN in tissues,methylation specific PCR was used to detect PTEN methylation levels,and Pearson correlation analysis was used to analyze the correlation between miR-4465 and PTEN in cancer tissues,and the relationships between PTEN gene promoter methylation rate and gender,age,clinical classification,tumor staging,and lymph node metastasis.Hep-2 cells were divided into blank group,control group(transfected with mimic NC),and miR-4465 mimic group(transfected with miR-4465 mimic).PTEN mRNA expression in Hep-2 cells was detected by qRT-PCR,PTEN protein expression in Hep-2 cells was detected by Western blot,the proliferation of Hep-2 cells was detected by CCK-8,the invasion of Hep-2 cells was detected by Transwell assay,and the migration of Hep-2 cells was detected by cell scratch healing assay.Hep-2 cells were further divided into}group(transfected with pcDNA3.0 plasmid)and pcDNA3.0-PTEN group(transfected with pcDNA3.0-PTEN plasmid).Cell proliferation was detected by CCK-8,cell invasion was detected by Transwell assay,and cell migration was detected by cell scratch healing assay.Results The relative expressions of miR-4465 and PTEN in laryngeal carcinoma tissues were lower than those in adjacent tissues(P<0.001).The expression of miR-4465 in cancer tissue was positively correlated with PTEN(r=0.459,P=0.036).The methylation rate of PTEN in cancerous tissue was significantly higher than that in paracancerous tissue(P<0.001).There was no correlation between the methylation rate of

关 键 词：喉癌 miR-4465 PTEN甲基化 增殖 侵袭 迁移 

分 类 号：R739.65[医药卫生—肿瘤]