抑制miR-182表达对裸鼠乳腺癌皮下移植瘤生长和转移的影响及其机制  

Effect of inhibiting expression of miR-182 on growth and metastasis of breast cancer xenografts in nude mice and its mechanism

作　　者：董超 黄远丽 徐正丰 DONG Chao;HUANG Yuanli;XU Zhengfeng(Jingzhou Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology,Jingzhou 434020,China)

机构地区：[1]华中科技大学同济医学院附属荆州医院

出　　处：《山东医药》2019年第31期42-45,50,共5页Shandong Medical Journal

摘　　要：目的探讨抑制miR-182表达对裸鼠乳腺癌皮下移植瘤生长和转移的影响及其机制。方法通过脂质体转染技术,将miR-182抑制物(miR-182 inhibitor)及其阴性对照物(inhibitor NC)转染至乳腺癌MDA-MB-231细胞,构建抑制miR-182表达的MDA-MB-231细胞和阴性对照MDA-MB-231细胞。选择雌性BALB/c裸鼠20只,随机分为观察组、对照组各10只,分别于右侧后腿皮下接种转染miR-182 inhibitor、inhibitor NC的MDA-MB-231细胞,观察皮下移植瘤生长情况。接种30 d处死,取出裸鼠皮下移植瘤组织和两侧腹股沟淋巴结,称取皮下移植瘤质量,采用苏木素-伊红染色观察腹股沟淋巴结,计算淋巴结转移率和阳性率。采用RT-qPCR法检测皮下移植瘤组织miR-182表达,采用Western blotting法检测皮下移植瘤组织叉头框转录因子O亚族1(FOXO1)、Bcl-2、Bax蛋白表达。结果转染miR-182 inhibitor的MDA-MB-231细胞miR-182相对表达量明显低于转染inhibitor NC的MDA-MB-231细胞(P<0.01)。两组均成功构建皮下移植瘤模型,无死亡。观察组皮下移植瘤生长速度和瘤体质量均低于对照组,并且腹股沟淋巴结转移率和阳性率均低于对照组(P均<0.05)。与对照组比较,观察组皮下移植瘤组织miR-182相对表达量低于对照组(P<0.05)。观察组皮下移植瘤组织FOXO1、Bax蛋白相对表达量低于对照组,Bcl-2蛋白相对表达量高于对照组(P均<0.05)。结论抑制miR-182表达可能通过下调靶基因FOXO1表达来抑制裸鼠乳腺癌皮下移植瘤的生长和转移。Objective To investigate the effects of inhibiting the expression of miR-182 on the growth and metastasis of breast cancer subcutaneously transplanted tumors in nude mice and its mechanism.Methods The miR-182 inhibitor and inhibitor negative control(NC)were transfected into breast cancer MDA-MB-231 cells by liposome transfection technique.MDA-MB-231 cells with inhibited expression of miR-182,and MDA-MB-231 cells as the negative controls were constructed.Twenty female BALB/c nude mice were randomly divided into the observation group and control group with 10 mice in each group.MDA-MB-231 cells transfected with miR-182 inhibitor and inhibitor NC were inoculated subcutaneously on the right hind leg to observe the growth of subcutaneous transplanted tumors.Thirty days after inoculation,the nude mice were sacrificed.Subcutaneous transplanted tumor tissues and bilateral inguinal lymph nodes were taken out.The quality of subcutaneous transplanted tumors was evaluated.The inguinal lymph nodes were observed by hematoxylin-eosin staining.The lymph node metastasis rate and positive rate were calculated.The expression of miR-182 was detected by RT-qPCR,and the expression of FOXO1,Bcl-2 and Bax was detected by Western blotting.Results The relative expression of miR-182 in MDA-MB-231 cells transfected with miR-182 inhibitor was significantly lower than that in MDA-MB-231 cells transfected with inhibitor NC(P<0.01).Subcutaneous transplantation models were successfully established in both groups,and no death occurred.The growth rate and mass of subcutaneous transplanted tumors in the observation group were lower than those in the control group,and the rate of inguinal lymph node metastasis and positive rate were lower than those in the control group(all P<0.05).Compared with the control group,the relative expression of miR-182 in the subcutaneous transplanted tumors in the observation group was lower than that in the control group(P<0.05).The relative expression of FOXO1 and Bax protein in subcutaneous transplanted tumors of the

关 键 词：乳腺癌 移植瘤 微小RNA-182 叉头框转录因子O亚族1 生长 转移 裸鼠 

分 类 号：R737.9[医药卫生—肿瘤]