MiR-651-3p调控乳腺癌细胞中免疫抑制因子IL-10与TGFβ1表达的机制研究  

miR-651-3p suppresses the expression of immunosuppressive factors IL-10 and TGFβ1 in breast cancer

作　　者：王珏 何欣 徐忠玲 肖铟 WANG Jue;HE Xin;XU Zhongling;XIAO Yin(Department of Oncology,Third Affiliated Hospital of Qiqihar Medical College,Qiqihaer 161000,China)

机构地区：[1]齐齐哈尔医学院附属第三医院肿瘤一科,161000

出　　处：《免疫学杂志》2023年第12期1050-1057,共8页Immunological Journal

基　　金：齐齐哈尔市科技计划联合引导项目(LSFGG-2022009)

摘　　要：目的探讨miR-651-3p在乳腺癌中的表达以及在调控免疫抑制因子IL-10与TGFβ1的表达和乳腺癌细胞凋亡水平中的作用。方法qRT-PCR检测miR-651-3p的表达水平,应用过表达与沉默miR-651-3p的质粒以及敲减SP2表达的质粒转染乳腺癌MCF-7细胞,Western blot检测SP2、IL-10与TGFβ1的表达水平变化,流式细胞仪检测MCF-7细胞凋亡水平变化,双荧光素酶报告基因验证miR-651-3p与SP2的靶向结合,ChIP实验验证SP2分别与IL-10、TGFβ1启动子区的结合作用。结果miR-651-3p在乳腺癌细胞中低表达(P<0.05),单独过表达miR-651-3p与单独敲低SP2的表达显著下调IL-10与TGFβ1的表达水平,促进乳腺癌细胞凋亡水平,而沉默miR-651-3p作用则相反(P<0.05)。miR-651-3p与SP23’-UTR端结合并下调其表达(P<0.05),在沉默miR-651-3p基础上沉默SP2可显著减弱沉默miR-651-3p对乳腺癌细胞中IL-10与TGFβ1的表达以及凋亡水平的影响(P<0.05)。SP2通过分别结合IL-10与TGFβ1的启动子区促进二者表达水平。结论在乳腺癌中低表达的miR-651-3p通过靶向抑制SP2的表达,进而减少SP2对IL-10与TGFβ1表达的上调作用,抑制乳腺癌细胞中免疫抑制因子IL-10与TGFβ1的表达并促进乳腺癌细胞凋亡。This study was aimed to investigate the expression of miR-651-3p in breast cancer and its role in regulating the expression of immunosuppressive factors IL-10 and TGFβ1 as well as the apoptosis level of breast cancer cells.qRT-PCR was used to detect the expression level of miR-651-3p in MCF-7 cells.miR-651-3p-over-expressng/-silencing plasmids and SP2-silencing plasmid were used to transfect MCF-7 cells.The expression levels of SP2,IL-10 and TGFβ1 were detected by Western blot assays.The apoptosis level of MCF-7 cells was detected by flow cytometry.The binding between miR-651-3p and SP2 was verified by dual luciferase gene report,and the binding of SP2 to IL-10 and TGFβ1 promoter region was verified by ChIP assay.Our results showed that miR-651-3p was down-regulated in breast cancer cells(P<0.05).The over-expression of miR-651-3p and the knockdown of SP2 significantly down-regulated the expression levels of IL-10 and TGFβ1 and promoted the apoptosis of breast cancer cells,while silenced miR-651-3p had the opposite effect.miR-651-3p binds to the SP2-3’-UTR end and down-regulates its expression.Based on the silencing of miR-651-3p,the silencing of SP2 can significantly reduce the influence of silenced miR-651-3p on the expression of IL-10 and TGFβ1 and apoptosis level in breast cancer cells.SP2 promotes the expression levels of IL-10 and TGFβ1 by binding to their promoters,respectively.Taken together,down-regulated miR-651-3p in breast cancer inhibited the expression of SP2 by binding to its 3’-UTR region,reduced the up-regulation effect of SP2 on the expression of IL-10 and TGFβ1,thus inhibiting the expression of immunosuppressive factors IL-10 and TGFβ1 and promoting apoptosis in breast cancer cells.

关 键 词：miR-651-3p SP2 免疫功能 乳腺癌 

分 类 号：R739.9[医药卫生—肿瘤]