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南京医科大学学报(自然科学版)                                 第45卷第12期
               ·1766 ·                    Journal of Nanjing Medical University(Natural Sciences)  2025年12月


             ·基础研究·

              RBM7在乳腺癌细胞MDA⁃MB⁃231中的表达及其对AKAP12表

              达的影响



              石   靓 ,奚佩雯 ,吴        靓 ,张 旭 ,丁       强  1*
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               南京医科大学第一附属医院乳腺中心,健康管理中心,病理科,江苏                        南京 210029
              1                             2            3
             [摘    要] 目的:探讨RNA结合蛋白7(RNA binding motif protein 7,RBM7)在人源性乳腺癌细胞MDA⁃MB⁃231中的表达,及其
              对A激酶锚定蛋白12(A⁃kinase anchoring protein 12,AKAP12)表达的影响。方法:在人源性乳腺癌细胞MDA⁃MB⁃231中分别转
              染RBM7过表达、干扰慢病毒(实验组)和相应对照慢病毒(对照组),经嘌呤霉素筛选后获得稳定转染的细胞株,使用荧光显微
              镜观察细胞转染情况。通过qRT⁃PCR和Western blot实验分别验证转染后细胞内RBM7的表达情况,并观察RBM7表达改变对
              AKAP12表达的影响。经RNA测序(RNA⁃sequence,RNA⁃seq)获得的差异表达基因进行基因本体论(gene ontology,GO)富集分
              析发现受RBM7影响的差异表达基因的主要富集通路,并结合UALCAN数据库分析AKAP12在乳腺癌中的表达情况。更进一
              步通过 RNA 结合蛋白免疫共沉淀(RNA binding protein immunoprecipitation,RIP)实验明确 RBM7 与 AKAP12 之间的关系。通
              过免疫组化在乳腺癌组织中明确 RBM7 表达与 AKAP12 表达之间的关系。结果:在乳腺癌细胞 MDA⁃MB⁃231 中分别转染
              RBM7过表达、干扰慢病毒,经嘌呤霉素筛选2周后,获得RBM7过表达及干扰的稳转细胞株。经RNA⁃seq获得的差异表达基
              因进行GO富集分析发现,受RBM7显著影响的基因主要富集在细胞周期通路,且UALCAN数据库分析发现AKAP12在乳腺癌
              中低表达(P < 0.05)。qRT⁃PCR和Western blot显示,RBM7过表达能下调AKAP12的RNA及蛋白的表达(P < 0.05),RBM7干扰
              后能上调AKAP12的RNA及蛋白表达(P < 0.05)。RIP实验显示,RBM7能直接结合AKAP12 mRNA从而发挥其对AKAP12的
              调节作用(P < 0.05)。免疫组化结果表明,在人乳腺癌组织中RBM7表达与AKAP12表达呈显著负性关联(P < 0.05)。结论:
              RBM7在乳腺癌细胞(MDA⁃MB⁃231)和乳腺癌组织中下调AKAP12的表达。
             [关键词] 乳腺癌;RNA结合蛋白7;A激酶锚定蛋白12;细胞周期
             [中图分类号] R737.9                   [文献标志码] A                      [文章编号] 1007⁃4368(2025)12⁃1766⁃09
              doi:10.7655/NYDXBNSN250949


              The expression of RBM7 in breast cancer cell MDA⁃MB⁃231 and its effects on AKAP12
              expression

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              SHI Liang ,XI Peiwen ,WU Jing ,ZHANG Xu ,DING Qiang 1*
               Breast Disease Center,Health Management Center,Department of Pathology,the First Affiliated Hospital of
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              Nanjing Medical University,Nanjing 210029,China
             [Abstract] Objective: To investigate the expression of RNA binding motif protein 7(RBM7)in human breast cancer cell MDA⁃MB⁃
              231,and its effects on the expression of A⁃kinase anchoring protein 12(AKAP12). Methods:MDA⁃MB⁃231 cells were transfected with
              lentiviruses for RBM7 overexpression,knockdown(experimental group)and the corresponding control lentivirus(control group)
              respectively. Stable transfected cell lines were selected with puromycin and verified via fluorescence microscopy. The expression of
              RBM7 after transfection was verified by qRT⁃PCR and Western blot experiments,respectively,and the effect of RBM7 expression
              change on AKAP12 expression was observed. Gene ontology(GO)enrichment analysis was performed on the differentially expressed
              genes obtained by RNA⁃sequence,and revealed significantly enriched pathways regulated by RBM7. At the same time,the UALCAN
              database was employed to assess AKAP12 expression in breast cancer. The relationship between RBM7 and AKAP12 was studied by
              RNA binding protein immunoprecipitation(RIP)assays. Furthermore,immunohistochemicalanalysis was performed to delineate the


             [基金项目] 国家自然科学基金(82372770,82403190)
              通信作者(Corresponding author),E⁃mail: dingqiang@njmu.edu.cn(ORCID:0000⁃0003⁃1499⁃6200)
              ∗
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