HMGB2沉默对乳腺癌MDA-MB-231细胞增殖、凋亡及侵袭迁移的影响  

Effects of HMGB2 silencing on proliferation,apoptosis,invasion and migration of breast cancer cell line MDA-MB-231

作　　者：刘爱蕙[1] 李秀楠[1] 王钢乐[1] 唐欣[1] 董懿[1] 纪楠[1] 康骅[2] LIU Aihui LI Xiunan WANG Gangyue TANG Xin DONG Yi JI Nan KANG Hua(Department of Breast Oncology, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing 100006, China)

机构地区：[1]首都医科大学附属妇产医院乳腺科,北京100006 [2]首都医科大学宣武医院普外科,100050

出　　处：《临床肿瘤学杂志》2017年第1期1-5,共5页Chinese Clinical Oncology

基　　金：首都医科大学附属北京妇产医院中青年学科骨干培养专项资助项目(fcyy201530)

摘　　要：目的探讨HMGB2沉默对乳腺癌MDA-MB-231细胞增殖、凋亡及侵袭迁移的影响。方法采用将靶向HMGB2的寡核苷酸克隆至p LKO.1 puro载体质粒构建靶向沉默HMGB2表达的慢病毒shRNA。将MDA-MB-231细胞分为对照组、空转染组及HMGB2干扰组,其中空转染组及HMGB2干扰组分别转染包含随机序列shRNA和靶向HMGB2 shRNA的病毒液,对照组仅行常规培养而不行任何转染。采用Western blotting检测各组转染48 h后的HMGB2蛋白水平以评价转染效率,采用MTT法计算各组细胞的存活率,流式细胞术检测转染48 h后的细胞凋亡率,Transwell法检测转染48 h后的细胞侵袭和转移水平。结果 HMGB2干扰组的HMGB2蛋白水平低于对照组和空转染组,差异有统计学意义(P<0.05),提示构建的载体沉默HMGB2的效果显著。与对照组和空转染组比较,HMGB2干扰组的细胞存活率降低,且随着转染时间的延长而呈降低趋势,差异均有统计学意义(P<0.05);HMGB2干扰组细胞的早期、晚期及总凋亡率均高于其余两组,且侵袭细胞数目和迁移细胞数目均低于其余两组,以上差异均有统计学意义(P<0.05)。结论 shRNA靶向沉默MDA-MB-231细胞中HMGB2表达效果显著,可抑制细胞增殖及侵袭迁移并诱导细胞凋亡,为乳腺癌靶向治疗提供一定参考。Objective To investigate the effect of HMGB2 silencing on proliferation, apoptosis, invasion and migration of breast cancer cell line MDA-MB-231. Methods The lentiviral vector was constructed by cloning the oligonucleotides targeting HMGB2 into pLKO. 1 puro vector. MDA-MB-231 cells were divided into control group, empty transfection group and HMGB2 interference group. The empty transfection group was transfected with random sequence shRNA and HMGB2 interference group was transfected with lentivi- ral vector targeting HMGB2 shRNA. The control group was treated with routine culture without any transfection. Western blotting was used to detect the level of HMGB2 protein after transfection for 48 h in each group to evaluate the efficiency of transfection. MTY meth- od was used to calculate the survival rate of each group. Flow cytometry was used to detect the apoptosis rate after transfection for 48 h. The Transwell method was used to detect the invasion and metastasis of 48 h after transfection. Results The level of HMGB2 protein in the HMGB2 interference group was lower than those in control group and empty transfection group, and the difference was statistical- ly significant （P〈0. 05）. Compared with control group and empty transfection group, the survival rate of HMGB2 interference group was decreased, and there was a decreasing trend with the increase of transfection time （P〈0.05）. The early, late and total apoptosis rates of HMGB2 interference group were higher than those in the other two groups, and the number of invasive ceils and the number of migrating ceils were lower than the other two groups （P〈0.05）. Conclusion Silencing HMGB2 expression via shRNA in MDA-MB-231 cells has a significant effect, can inhibit cell proliferation, invasion, migration and induce cell apoptosis, to provide a reference for breast cancer targeted therapy.

关 键 词：乳腺癌 高迁移率族蛋白2(HMGB2) 增殖 凋亡 侵袭 迁移 

分 类 号：R737.9[医药卫生—肿瘤]