Hsa-miR-212靶向调控BRCA1对乳腺癌MDA-MB-231细胞放疗敏感性的影响  

Effect of Hsa-miR-212 targeting BRCA1 on the radio sensitivity of breast cancer MDA-MB-231 cells

作　　者：尹婷婷[1] 王清云 胡爽 王秀艳[1] YIN Ting-ting;WANG Qing-yun;HU Shuang;WANG Xiu-yan(First Affiliated Hospital of Xingtai Medical College,Xingtai First Hospital,Laboratory Department;First Affiliated Hospital of Xingtai Medical College,Xingtai First Hospital,Head,Neck and Breast Department,Xingtai 054001,China)

机构地区：[1]邢台医学高等专科学校第一附属医院(邢台市第一医院)检验科,河北邢台054001 [2]邢台医学高等专科学校第一附属医院(邢台市第一医院)头颈乳腺科,河北邢台054001

出　　处：《生物技术》2022年第5期636-641,629,共7页Biotechnology

基　　金：邢台市重点研发计划项目(2020ZC198)。

摘　　要：[目的]探讨Hsa-miR-212靶向调控BRCA1对乳腺癌MDA-MB-231细胞放疗敏感性的影响。[方法]设MDA-MB-231细胞组、RT组(X射线以12Gy/min的剂量率发射,照射孵育时间为48h)、Hsa-miR-212 mimics组、RT+Hsa-miR-212 mimics组,各组设6个平行样,培养72 h。试验结束后,采用细胞计数试剂盒-8和结晶紫测定增殖及单克隆形成数目,流式细胞仪测量细胞凋亡水平,RT-PCR法及蛋白印迹法测定Hsa-miR-212、BRCA1 mRNA和蛋白水平。[结果]与MDA-MB-231细胞组比较,RT组、RT+Hsa-miR-212 mimics组OD值、存活率、细胞克隆形成数目、Hsa-miR-212 mRNA、BRCA1 mRNA与蛋白、穿膜数均降低(P<0.05),Hsa-miR-212 mimics组OD值、存活率、细胞克隆形成数目、Hsa-miR-212 mRNA、BRCA1 mRNA与蛋白、穿膜数升高(P<0.05);与RT组比较,Hsa-miR-212 mimics组、RT+Hsa-miR-212 mimics组OD值、存活率、细胞克隆形成数目、Hsa-miR-212 mRNA、BRCA1 mRNA与蛋白、穿膜数升高(P<0.05);与Hsa-miR-212 mimics组比较,RT+Hsa-miR-212 mimics组OD值、存活率、细胞克隆形成数目、Hsa-miR-212 mRNA、BRCA1 mRNA与蛋白、穿膜数降低(P<0.05)。与MDA-MB-231细胞组比较,RT组、RT+Hsa-miR-212mimics组凋亡率升高(P<0.05),Hsa-miR-212 mimics组凋亡率降低、穿膜数升高(P<0.05);与RT组比较,Hsa-miR-212 mimics组、RT+Hsa-miR-212 mimics组凋亡率降低(P<0.05);与Hsa-miR-212 mimics组比较,RT+Hsa-miR-212 mimics组凋亡率升高、穿膜数降低(P<0.05)。[结论]Hsa-miR-212过表达通过靶向BRCA1 mRNA的3'非编码区,降低BRCA1的翻译水平,并降低了乳腺癌细胞MDA-MB-231的放射敏感性。[Objective]To investigate the effect of Hsa-miR-212 targeting BRCA1 on the radiosensitivity of breast cancer MDA-MB-231 cells.[Method]MDA-MB-231 cell group,RT group(X-ray generator emits at a dose rate of 12Gy/min for 48 hours),Hsa-miR-212 mimics group,RT+Hsa-miR-212 mimics group were set up with 6 parallel samples in each group,and cultured for 72h.After the experiment,cell counting kit-8 and crystal violet were used to determine the number of proliferation and monoclonal formation,flow cytometry used to measure the level of cell apoptosis,and RT-PCR and Western Blotting were used to determine Hsa-miR-212,BRCA1 mRNA and protein level.[Result]Compared with the MDA-MB-231 cell group,the OD value,survival rate,cell clone formation number,Hsa-miR-212 mRNA,BRCA1 mRNA and protein,and the number of membrane penetration in the RT group and RT+Hsa-miR-212 mimics group were all decreased(P<0.05),and the OD value,survival rate,cell clone formation number,Hsa-miR-212 mRNA,BRCA1 mRNA and protein,the number of transmembrane in the Hsa-miR-212 mimics group increased(P<0.05).Compared with the RT group,the OD value,survival rate,cell clone formation number,Hsa-miR-212 mRNA,BRCA1 mRNA and protein,the number of membrane penetration in the Hsa-miR-212 mimics group and the RT+Hsa-miR-212 mimics group increased(P<0.05).Compared with Hsa-miR-212 mimics group,the OD value,survival rate,cell clone formation number,Hsa-miR-212 mRNA,BRCA1 mRNA and protein,and membrane penetration in the RT+Hsa-miR-212 mimics group decreased(P<0.05).Compared with the MDA-MB-231 cell group,the apoptosis rate in the RT group and RT+Hsa-miR-212 mimics group increased(P<0.05),and the apoptosis rate in the Hsa-miR-212 mimics group decreased(P<0.05),and compared with the RT group,the apoptosis rate of the Hsa-miR-212 mimics group and the RT+Hsa-miR-212 mimics group decreased(P<0.05).Compared with the Hsa-miR-212 mimics group,the apoptotic rate in the RT+Hsa-miR-212 mimics group increased(P<0.05).[Conclusion]Hsa-miR-212 overexpression reduces the translation leve

关 键 词：Hsa-miR-212 BRCA1 乳腺癌 放疗敏感性 

分 类 号：R737.9[医药卫生—肿瘤]