mRNA稳定因子HuR通过cyclinD1的靶向作用对人乳腺癌MCF-7细胞株行为的影响  

Effect of mRNA stability factor HuR on biological behavior of human breast cancer cells through targeting cyclinD1

作　　者：郑克思 郑翔 曾勇 陈聪 吴元肇 郑克文[3] ZHENG Kesi;ZHENG Xiang;ZENG Yong(Department of Oncology,Wenzhou People’s Hospital,the Third Clinical College of Wenzhou Medical University,Wenzhou 325000,China)

机构地区：[1]温州市人民医院、温州医科大学温州市第三临床学院肿瘤外科,325000 [2]温州市人民医院、温州医科大学温州市第三临床学院病理科,325000 [3]温州医科大学附属第一医院、温州医科大学第一临床学院泌尿外科

出　　处：《浙江医学》2019年第11期1128-1132,1157,共6页Zhejiang Medical Journal

基　　金：浙江省自然科学基金(LQ17H050002);温州市公益性科技计划项目(Y20160336、Y20170752)

摘　　要：目的探究mRNA稳定因子人抗原R(human antigen R,HuR)通过细胞周期特异性蛋白D1(cyclinD1)对人乳腺癌MCF-7细胞株增殖、凋亡和迁移等生命活动的影响。方法将MCF-7细胞分为对照组、HuR过表达组(构建HuR过表达载体并转染)、Hu R敲低组(构建HuR敲低载体并转染)。各组细胞分别进行MTT检测细胞增殖活性;Annexin V-APC/7-AAD双染检测细胞凋亡;Transwell实验检测细胞迁移能力;荧光定量PCR及Western blot检测HuR、cyclinD1的表达。结果与对照组比较,HuR过表达组细胞的增殖率、迁移细胞数量显著上升(t=-18.29、-4.57,P<0.01或0.05),细胞凋亡率显著降低(t=3.70,P<0.05),HuR(t=-11.70、-10.55,均P<0.01)、cyclinD1(t=-9.34、-6.75,均P<0.01)的m RNA及蛋白表达显著升高;HuR敲低组细胞的增殖率、迁移细胞数量显著下降(t=13.83、3.91,P<0.05或0.01),细胞凋亡率显著升高(t=-45.32,P<0.01),HuR(t=21.88、21.35,均P<0.01)、cyclinD1(t=9.55、4.33,P<0.01或0.05)的m RNA及蛋白表达显著降低。结论HuR可能通过cyclinD1促进人乳腺癌MCF-7细胞的增殖、迁移,抑制细胞的凋亡,可能为靶向治疗乳腺癌提供新的潜在靶标。Objective To investigate the effects of mRNA stability factor human antigen R(HuR)on proliferation,apoptosis and migration of human breast cancercells and its relation to cyclinD1.Methods Human breast cancerMCF-7 cells were transfected withHuR overexpression vector(overExp-HuR group)orHuR knockdown vector(cas9-HuRgroup).The cell proliferation was detected by MTT,the apoptosis was detected by Annexin V-APC/7-AAD double staining,the cell migration was detected by Transwell assay,and the expression of Hu R and cyclinD1 were detected by fluorescence quantitative PCR and Western blot.Results Compared with the control group,the proliferation rateand the number of migrating cells of overExp-HuR group increased significantly(t=-18.29,P<0.01;t=-4.57,P<0.05),the apoptosis rate of overExp-HuR group decreased(t=3.70,P<0.05),the m RNA and protein expression of HuR(t=-11.70,-10.55,P<0.01)and cyclinD1(t=-9.34,-6.75,P<0.01)of overExp-HuR group increased significantly.The proliferation rateand the number of migrating cellsincas9-HuR group decreased significantly(t=13.83,P<0.01;t=3.91,P<0.05),and the apoptosis rateincreased significantly(t=-45.32,P<0.01),the m RNA and protein expression of HuR(t=21.88,21.35,P<0.01)and cyclinD1(t=9.55,4.33,P<0.01 or 0.05)in cas9-HuRdecreased significantly.Conclusion HuR can promote the proliferation and migration,inhibit the apoptosisof human breast cancer MCF-7 cells through cyclinD1,which may provide a potential target for treatment of breast cancer.

关 键 词：人抗原R 细胞周期特异性蛋白D1 迁移 增殖 凋亡 

分 类 号：R737.9[医药卫生—肿瘤]