hsa_circ_0000520通过调控miR-556-5p/SLC38A2促进乳腺癌的发生和转移  

hsa_circ_0000520 promotes occurrence and metastasis of breast cancer by regulating miR-556-5p/SLC38A2

作　　者：张景臣[1] 李新[1] 李江涛[1] 李海平[1] 陈艳丽 牛冰[1] 祁川川 叶贝贝 ZHANG Jingchen;LI Xin;LI Jiangtao;LI Haiping;CHEN Yanli;NIU Bing;QI Chuanchuan;YE Beibei(Department of Galactophore,Zhengzhou People's Hospital,Henan Zhengzhou 450053,China)

机构地区：[1]郑州人民医院乳腺科,河南郑州450053

出　　处：《现代肿瘤医学》2023年第12期2190-2196,共7页Journal of Modern Oncology

基　　金：河南省医学科技攻关计划(联合共建)项目(编号:LHGJ20191074,201303219)。

摘　　要：目的:探究hsa_circ_0000520促进乳腺癌发生和转移的作用机制。方法:双荧光素酶报告基因实验、RNA pull-down实验验证miR-556-5p与hsa_circ_0000520、溶质载体家族38成员2(SLC38A2)的靶向关系。MCF7细胞分为sh-NC组、sh-hsa_circ_0000520组、sh-hsa_circ_0000520+anti-NC组、sh-hsa_circ_0000520+anti-miR-556-5p组,qRT-PCR或Western blot检测细胞中hsa_circ_0000520、miR-556-5p、SLC38A2表达水平;MTT检测、平板克隆形成实验评估细胞增殖能力;划痕愈合实验、Transwell实验评估细胞迁移、侵袭能力。通过裸鼠成瘤实验评估hsa_circ_0000520对miR-556-5p/SLC38A2的调控作用及对移植瘤生长的影响。结果:经验证,MCF7细胞中miR-556-5p与hsa_circ_0000520、SLC38A2均存在靶向关系。与sh-NC组比较,sh-hsa_circ_0000520组可降低MCF7细胞中hsa_circ_0000520、SLC38A2 mRNA和蛋白表达水平、细胞活力、克隆形成数目、划痕愈合率及迁移、侵袭细胞数(P<0.05),升高miR-556-5p表达水平(P<0.05);与sh-hsa_circ_0000520+anti-NC组比较,sh-hsa_circ_0000520+anti-miR-556-5p组可降低MCF7细胞中miR-556-5p表达水平(P<0.05),升高SLC38A2 mRNA和蛋白表达水平、细胞活力、克隆形成数目、划痕愈合率及迁移、侵袭细胞数(P<0.05),而对hsa_circ_0000520表达无显著影响(P>0.05)。裸鼠成瘤实验结果表明,敲低移植瘤中hsa_circ_0000520的表达可升高miR-556-5p表达水平并降低SLC38A2 mRNA和蛋白表达水平,同时降低肿瘤体积和肿瘤重量(P<0.05)。结论:hsa_circ_0000520可能通过靶向调控miR-556-5p/SLC38A2促进乳腺癌的发生和转移。Objective:To explore the mechanism of hsa_circ_0000520 in promoting the occurrence and metastasis of breast cancer.Methods:Dual-luciferase reporter gene experiments and RNA pull-down experiments verified the targeting relationship of miR-556-5p with hsa_circ_0000520 and solute carrier family 38 member 2(SLC38A2).MCF7 cells were separated into sh-NC group,sh-hsa_circ_0000520 group,sh-hsa_circ_0000520+anti-NC group,and sh-hsa_circ_0000520+anti-miR-556-5p group.qRT-PCR or Western blot were applied to detect the expression levels of hsa_circ_0000520,miR-556-5p and SLC38A2 in cells.MTT assay and plate colony formation assay were used to evaluate cell proliferation.Scratch-healing assay and Transwell assay were used to evaluate cell migration and invasion abilities.The nude mice tumorigenesis experiments were applied to evaluate the regulatory effect of hsa_circ_0000520 on miR-556-5p/SLC38A2 and its effect on the growth of xenografts.Results:It had been verified that miR-556-5p had a targeting relationship with hsa_circ_0000520 and SLC38A2 in MCF7 cells.Compared with the sh-NC group,the sh-hsa_circ_0000520 group could reduce the expression levels of hsa_circ_0000520 and SLC38A2 mRNA and protein,cell viability,number of colonies,wound healing rate,and the numbers of migrating and invasive cells in MCF7 cells(P<0.05),increase the expression level of miR-556-5p(P<0.05).Compared with sh-hsa_circ_0000520+anti-NC group,sh-hsa_circ_0000520+anti-miR-556-5p group could reduce the expression level of miR-556-5p in MCF7 cells(P<0.05),increase the expression levels of SLC38A2 mRNA and protein,cell viability,number of colonies,wound healing rate,and numbers of migrating and invasive cells(P<0.05),however,it had no obvious effect on the expression of hsa_circ_0000520(P>0.05).The results of tumorigenesis experiments in nude mice showed that knocking down the expression of hsa_circ_0000520 in xenograft tumors could increase the expression level of miR-556-5p and decrease the expression levels of SLC38A2 mRNA and protein,and reduce tum

关 键 词：hsa_circ_0000520 miR-556-5p 溶质载体家族38成员2 乳腺癌 

分 类 号：R737.9[医药卫生—肿瘤]