MiR-6838-5p过表达下调DDR1基因表达抑制乳腺癌MCF-7细胞的增殖  

MiR-6838-5p overexpression inhibits proliferation of breast cancer MCF-7 cells bydownregulating DDR1 expression

作　　者：薛良军 谈秋瑜 许静文 冯璐 李文锦 颜亮 李玉磊 XUE Liangjun;TAN Qiuyu;XU Jingwen;FENG Lu;LI Wenjin;YAN Liang;LI Yulei(Department of Radiotherapy,First Affiliated Hospital of Wannan Medical College,Wuhu 241002,China;School of Basic Medicine,Wannan Medical College,Wuhu 241002,China)

机构地区：[1]皖南医学院第一附属医院放疗科,安徽芜湖241002 [2]皖南医学院基础医学院,安徽芜湖241002

出　　处：《南方医科大学学报》2024年第9期1677-1684,共8页Journal of Southern Medical University

基　　金：安徽省高校科研编制重大项目(2022AH040179);安徽省大学生创新创业训练计划项目(S202310368008,S202310368041)。

摘　　要：目的探索miR-6838-5p调控DDR1基因表达水平对乳腺癌MCF-7细胞增殖的影响。方法采用脂质体转染方法干预乳腺癌MCF-7细胞中miR-6838-5p及DDR1基因表达,设置组别如下:Control mimic;miR-6838-5p mimic;Control inhibitor;miR-6838-5p inhibitor;Control siRNA、DDR1 siRNA、Control vector、DDR1 vector及miR-6838-5p过表达+DDR1过表达共转染组。qRT-PCR实验检测miR-6838-5p在正常乳腺上皮细胞及乳腺癌细胞中的表达;通过TargetscanV 8.0等软件预测miR-6838-5p可能作用的靶基因;双荧光素酶报告基因实验验证miR-6838-5p与DDR1存在靶向结合位点;CCK-8实验及EdU实验检测细胞增殖情况;Western blotting实验检测DDR1表达情况;通过裸鼠荷瘤实验验证miR-6838-5p通过调控DDR1表达抑制了乳腺癌MCF-7细胞增殖。结果miR-6838-5p在乳腺癌细胞中表达量低于乳腺正常上皮细胞(P<0.05);对比对照组,miR-6838-5p能抑制乳腺癌细胞增殖(P<0.05);双荧光素酶报告基因实验证明miR-6838-5p可以靶向结合DDR1(P<0.01),Western blotting实验证明miR-6838-5p能调控DDR1基因表达;DDR1促进乳腺癌细胞的增殖(P<0.05);miR-6838-5p过表达+DDR1过表达共转染后能回复miR-6838-5p对乳腺癌细胞增殖的抑制作用(P<0.05);裸鼠荷瘤实验结果表明过表达miR-6838-5p后肿瘤体积明显减小,Western blotting结果表明肿瘤内miR-6838-5p过表达后抑制了DDR1的表达(P<0.05)。结论miR-6838-5p通过调控DDR1基因表达抑制了乳腺癌细胞的增殖。Objective To explore the regulatory effect of miR-6838-5p on DDR1 gene expression and proliferation of breast cancer cells.Methods The expression levels of miR-6838-5p in normal breast epithelial cells and breast cancer cells were detected using qRT-PCR,and the potential target genes of miR-6838-5p was predicted using TargetscanV 8.0.Double luciferase reporter gene experiment was performed to verify the binding between miR-6838-5p and DDR1.Breast cancer MCF-7 cells were transfected via liposome,miR-6838-5p mimic,miR-6838-5p inhibitor,DDR1 siRNA,DDR1-overexpresisng vector,or both miR-6838-5p mimic and DDR1-overexpressing vector,and the changes in cell proliferation were examined with CCK-8 and EdU assays;Western blotting was used to detect the expression of DDR1.The mediating role of DDR1 in miR-6838-5p overexpression-induced inhibition of MCF-7 cell proliferation was verified in a nude mouse model bearing MCF-7 cell xenografts.Results The expression of miR-6838-5p was significantly lower in breast cancer cells than in normal breast epithelial cells.In MCF-7 cells,miR-6838-5p overexpression induced significant inhibition of cell proliferation.Dual luciferase reporter gene experiment demonstrated a binding relationship between miR-6838-5p and DDR1(P<0.01).Western blotting showed that miR-6838-5p overexpression significantly lowered DDR1 expression in MCF-7 cells,and DDR1 overexpression promoted proliferation of the cells;co-transfection of the cells with DDR1-overexpressing vector significantly attenuated the inhibitory effect of miR-6838-5p mimic on cell proliferation.In the tumor-bearing nude mice,the xenografts overexpressing miR-6838-5p showed a significantly smaller volum with obviously the expression of DDR1.Conclusion Overexpression of miR-6838-5p inhibits breast cancer cell proliferation by regulating DDR1 expression.

关 键 词：miR-6838-5p 乳腺癌 DDR1 细胞增殖 

分 类 号：R737.9[医药卫生—肿瘤]