miR-346在乳腺癌中的表达及对细胞生长、侵袭和凋亡的影响  

Expression of microRNA-346 in breast carcinoma and its effect on cell proliferation,invasion and apoptosis

作　　者：李晶晶[1] 张文馨 张倩[1] 刘艳月[1] 谷红丽[1] 万菲菲 王晶[2] LI Jingjing;ZHANG Wenxin;ZHANG Qian;LIU Yanyue;GU Hongli;WAN Feifei;WANG Jing(Department of Infectious Diseases,First Affiliated Hospital of Zhengzhou University,Zhengzhou 450001,China;Department of Hepatobiliary Surgery,First Affi-liated Hospital of Zhengzhou University)

机构地区：[1]郑州大学第一附属医院感染科,郑州450001 [2]郑州大学第一附属医院肝胆外科

出　　处：《山西医科大学学报》2020年第12期1301-1307,共7页Journal of Shanxi Medical University

基　　金：河南省高等学校重点科研计划项目(18A320061);河南省重点研发与推广专项项目(1778)。

摘　　要：目的观察微小RNA(miRNA)-346对乳腺癌细胞增殖和侵袭能力的影响并探讨其相关调控机制。方法采用实时荧光定量PCR检测乳腺癌组织及其相应癌旁正常组织中miRNA-346的表达。采用实时荧光定量PCR检测乳腺癌细胞系(MCF-7,MDA-MB-231,MDA-MB-435,MDA-MB-468,T47D)与正常乳腺细胞(MCF-10A)中miRNA-346的表达。以miRNA-346表达最高的乳腺癌细胞系MDA-MB-468为对象,分别转染miRNA-346抑制剂和miR-NC,命名为miR-346抑制剂组和对照组。real-time PCR检测转染效果利用CCK-8试剂及Transwell小室分别检测转染后乳腺癌细胞增殖和侵袭转移能力。利用流式细胞仪检测miR-346表达对细胞凋亡的影响。生物信息学软件预测miR-346的靶基因。real-time PCR和Western blot检测乳腺癌细胞中靶基因的表达。结果乳腺癌组织中miR-346 mRNA的相对水平显著高于癌旁组织(P<0.01)。乳腺癌细胞系中miR-346 mRNA的相对水平均显著高于正常乳腺细胞,且以MDA-MB-468升高最明显(P<0.01)。miRNA-346抑制剂组miR-346的表达显著低于对照组(P<0.01)。与对照组比较,miR-346抑制剂组细胞凋亡率显著升高(P<0.01),迁移能力和侵袭能力明显降低(P<0.01)。生物信息学软件显示miR-346的靶基因是BRMS1。在乳腺癌组织中BRMS1 mRNA和蛋白相对表达量显著低于癌旁正常组织。与对照组比较,miR-346抑制剂组中BRMS1 mRNA表达显著增加,且miR-346与BRMS1 mRNA的表达水平呈负相关(r=-0.82,P<0.01)。结论miR-346在乳腺癌细胞中呈现高表达,体外实验发现miR-346低表达能够抑制乳腺癌细胞的增殖和侵袭能力,其机制可能与上调BRMS1有关。Objective To observe the effects of microRNA(miRNA)-346 on the proliferation and the invasion of breast cancer cells and to explore its potential regulatory mechanism.Methods The expression level of miR-346 in breast cancer tissues was detected by real-time PCR.The expression levels of miR-346 in breast cancer cell lines(MCF-7,MDA-MB-231,MDA-MB-435,MDA-MB-468,T47D)and normal breast cell line(MCF-10A)were detected by real-time PCR.The breast cancer cell line MDA-MB-468 with the highest miR-346 expression was used as the subject,and transfected with NC mimics(NC group)and miR-346 inhibitor(inhibitor group)using liposome,respectively.Real-time PCR was used to detect the transfection efficiency.Meanwhile,CCK-8 assay and Transwell assay were used to detect the effects of miR-346 on cell proliferation,migration and invasion.Flow cytometry was used to detect the apoptosis.Bioinformatics software was used to predict the target genes of miR-346,respectively.Real-time PCR and Western blot were used to detect the mRNA and protein levels of target genes in breast cancer cells.Results The miR-346 expression in breast cancer tissues was significantly higher than that in paracancerous tissues(P<0.01).The miR-346 expression in breast cancer cell lines was significantly higher than that of normal breast cell line,especially in MDA-MB-435(P<0.01).The expression of miR-346 in inhibitor group was significantly lower than that in miR-NC group(P<0.01).Compared with miR-NC group,the cell apoptosis was increased(P<0.01),the abilities of proliferation,migration and invasion in inhibitor group were significantly down-regulated(P<0.01).Bioinformatics software showed that the target gene of miR-346 is BRMS1.The BRMS1 mRNA and protein expression levels in breast cancer tissues were significantly higher than that in paracancerous tissues.Compared with miR-NC group,the BRMS1 mRNA expression was significantly increased in MDA-MB-468 cells in inhibitor group(P<0.01),and the expression of BRMS1 mRNA was negatively related with miR-346 expression(

关 键 词：乳腺癌 微小RNA-346 BRMS1 细胞侵袭 

分 类 号：R737.9[医药卫生—肿瘤]