核仁素诱导自噬促进乳腺癌细胞增殖的作用机制  

Nucleolin promotes breast cancer proliferation by enhancing cell autophagy

作　　者：李佳莉 杨萍 彭晨阳 任安琦 林之昊 潘柳齐 李安怡 武福云 LI Jiali;YANG Ping;PENG Chenyang;REN Anqi;LIN Zhihao;PAN Liuqi;LI Anyi;WU Fuyun(College of Basic Medicine,Hubei University of Medicine,Shiyan 442000,Hubei,China)

机构地区：[1]湖北医药学院基础医学院,十堰442000

出　　处：《医学研究与战创伤救治》2023年第1期7-12,共6页Journal of Medical Research & Combat Trauma Care

基　　金：国家自然科学基金(81702639);湖北医药学院大学生创新创业训练计划项目(S202010929002)。

摘　　要：目的核仁素Nucleolin(NCL)与多种肿瘤的发生发展密切相关,也是抗肿瘤的重要靶点。文中旨在研究NCL蛋白促进乳腺癌细胞增殖的机制,为乳腺癌的治疗提供新的靶点及治疗策略。方法NCL过表达实验分为4组:4TO组(细胞转染pcDNA/4T0空载体)、4TO-NCL组(细胞转染4TO-NCL过表达载体)、4TO+Rapamycin组(细胞转染pcDNA/4T0后加入雷帕霉素处理)、4TO-NCL+Rapamycin组(细胞转染4TO-NCL过表达载体后加入雷帕霉素处理)。NCL敲低表达实验也分为4组:siRNA组(细胞转染对照siRNA)、NCLsiRNA组(细胞转染NCL siRNA)、siRNA+Rapamycin组(细胞转染siRNA后加入雷帕霉素处理)、NCLsiRNA+Rapamycin组(细胞转染NCL siRNA后加入雷帕霉素处理)。分别检测NCL过表达和敲低表达对乳腺癌细胞自噬相关蛋白表达的影响。NCL对自噬小体形成的影响,实验分为4组:GFP组(转染带有绿色荧光蛋白标签的GFP空载体)、GFP-NCL组(转染带有绿色荧光的NCL融合蛋白的GFP-NCL质粒)、GFP+Rapamycin组(转染带有绿色荧光蛋白标签的GFP空载体24 h后,加入Rapamycin处理细胞12 h)、GFP-NCL+Rapamycin组(转染带有绿色荧光的NCL融合蛋白的GFP-NCL质粒24 h后,加入Rapamycin处理细胞12 h)。NCL过表达实验分为4组:pcDNA/4T0组(细胞转染pcDNA/4T0空载体)、pcDNA/4T0-NCL过表达组(细胞转染4TO-NCL过表达载体)、pcDNA/4T0+3-MA组(细胞转染pcDNA/4T0空载体质粒后加入3-MA处理)、pcDNA/4T0-NCL+3-MA组(细胞转染4TO-NCL过表达载体后加入3-MA处理)。NCL敲低表达实验也分为4组:siRNA组(细胞转染对照siRNA)、NCL siRNA组(细胞转染NCL siRNA)、siRNA+3-MA组(细胞转染siRNA后加入3-MA)、NCLsiRNA+3-MA组(细胞转染NCL siRNA后加入3-MA)。MTT实验、EdU实验及细胞划痕实验检测NCL对乳腺癌细胞增殖和迁移的影响。结果免疫印迹结果显示,4TO-NCL过表达组与4TO组相比,P62的表达降低,Beclin的表达水平明显升高。MTT结果显示,与4T0组细胞活力(103.60±2.88)相�Objective Breast cancer is a common malignant tumor in women,with high incidence rate and mortality.To study the mechanism of Nucleolin(NCL)protein in promoting the proliferation of breast cancer cells,and to provide new targets and therapeutic strategies for breast cancer treatment.Methods The NCL overexpressed cells were divided into four groups:pcDNA/4TO group(cells transfected with pcDNA/4T0 empty vector),pcDNA/4TO-NCL group(cells transfected with pcDNA/4TO-NCL vector),pcDNA/4TO+Rapamycin group(cells transfected with pcDNA/4T0 were treated with rapamycin),pcDNA/4TO-NCL+Rapamycin group(cells transfected with pcDNA/4TO-NCL vector were treated with rapamycin).The NCL knockdown cells were also divided into four groups:siRNA group(cells transfected with control siRNA),NCLsiRNA group(cells transfected with NCL siRNA),siRNA+Rapamycin group(cells transfected with siRNA were treated with rapamycin),and NCLsiRNA+Rapamycin group(cells transfected with NCL siRNA were treated with rapamycin).The expression of autophagy related proteins in breast cancer cells were detected.NCL overexpressed cells were divided into four groups:GFP group(transfected with GFP vector),GFP-NCL group(transfected with GFP-NCL plasmid),GFP+Rapamycin group(transfected with GFP vector and treated with Rapamycin)GFP-NCL+Rapamycin group(transfected with GFP-NCL vector and treated with Rapamycin).MTT,Edu and cell scratch assays were used to detect the proliferation of breast cancer cells.NCL overexpressed cells were divided into four groups:pcDNA/4T0 group(transfected with pcDNA/4T0 vector),pcDNA/4T0-NCL group(transfected with pcDNA/4TO-NCL vector),pcDNA/4T0+3-MA group(cells transfected with pcDNA/4T0 vector and treated with 3-MA),pcDNA/4T0-NCL+3-MA group(cells transfected with pcDNA/4TO-NCL vector and treated with 3-MA).NCL knockdown cells were also divided into four groups:siRNA group(cells transfected with control siRNA),NCL siRNA group(cells transfected with NCL siRNA),siRNA+3-MA group(cells transfected with siRNA and treated with 3-MA),and NCL siR

关 键 词：核仁素 乳腺癌细胞 自噬 增殖 

分 类 号：R737.9[医药卫生—肿瘤]