中国癌症杂志 ›› 2020, Vol. 30 ›› Issue (12): 984-990.doi: 10.19401/j.cnki.1007-3639.2020.12.003

• 论著 • 上一篇    下一篇

miR-6775-3p在乳腺癌细胞中的表达及其对乳腺癌细胞生物学行为的影响

齐 正 1 ,薛 云 2 ,李宗良 3 ,张小冲 1 ,雷秋香 1 ,张 提 4 ,景立红 5 ,刘登湘 1   

  1. 1. 邢台市人民医院检验科,河北 邢台 054031 ;
    2. 邢台市第三医院检验科,河北 邢台 054031 ;
    3. 邢台市人民医院核医学科,河北 邢台 054031 ;
    4. 邢台市人民医院美容科,河北 邢台 054031 ;
    5. 巨鹿县卫生学校内科,河北 邢台 055250
  • 出版日期:2020-12-30 发布日期:2021-01-08
  • 通信作者: 刘登湘 E-mail: rmyy666@163.com
  • 基金资助:
    河北省重点研发计划项目(182777122D)。

Expression of miR-6775-3p in breast cancer cells and its effect on biological behavior of breast cancer cells 

QI Zheng 1 , XUE Yun 2 , LI Zongliang 3 , ZHANG Xiaochong 1 , LEI Qiuxiang 1 , ZHANG Ti 4 , JING Lihong 5 , LIU Dengxiang 1#br#   

  1. 1. Department of Clinical Laboratory, Xingtai City People’s Hospital, Xingtai 054031, Hebei Province, China; 2. Department of Clinical Laboratory, Xingtai City Third Hospital, Xingtai 054031, Hebei Province, China; 3. Department of Nuclear Medicine, Xingtai People’s Hospital, Xingtai 054031, Hebei Province, China; 4. Department of Cosmetics, Xingtai People’s Hospital, Xingtai 054031, Hebei Province; 5. Department of Internal Medicine, Julu County Health School, Xingtai 055250, Hebei Province, China
  • Online:2020-12-30 Published:2021-01-08
  • Contact: LIU Dengxiang E-mail: rmyy666@163.com

摘要: 背景与目的:微小RNA(microRNA,miRNA)与肿瘤的发生、发展过程密切相关。探讨miR-6775-3p在乳腺癌细胞系中的表达及其对乳腺癌细胞生物学行为的影响。方法:通过实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR)检测4种乳腺癌细胞系MDA-MB-231、MDA-MB-453、MDA-MB-468和BT-549中miR-6775-3p的表达水平,选取miR-6775-3p表达水平最低的乳腺癌细胞系过表达miR-6775-3p后,采用细胞计数试剂盒(cell counting kit-8,CCK-8)法检测细胞的增殖情况,同时采用transwell迁移和侵袭实验分别检测细胞迁移和侵袭能力的变化。通过RTFQ-PCR和蛋白[质]印迹法(Western blot)检测miR-6775-3p过表达的乳腺癌细胞系中细胞周期蛋白依赖性蛋白激酶4(cyclin-dependent protein kinase 4,CDK4)和CDK6,以及侵袭转移标志物基质金属蛋白酶(matrix metalloproteinase,MMP)17和MMP24 mRNA以及蛋白的表达变化。结果:RTFQ-PCR结果显示,乳腺癌细胞系MDA-MB-453中miR-6775-3p的表达最低,在MDA-MB-453细胞中转染miR-6775-3p mimics后,miR-6775-3p的表达水平明显升高(P<0.001)。CCK-8实验结果显示,MDA-MB-453细胞过表达miR-6775-3p后,细胞的增殖能力明显降低(P<0.01)。Transwell迁移和侵袭实验结果显示,MDA-MB-453细胞过表达miR-6775-3p后,细胞的迁移(P<0.001)和侵袭能力(P<0.01)明显降低。RTFQ-PCR和Western blot实验结果显示,CDK4、CDK6及MMP17、MMP24的mRNA表达和蛋白水平均显著降低(P<0.01)。结论:miR-6775-3p可能抑制乳腺癌细胞的增殖、迁移和侵袭能力。

关键词: miR-6775-3p, 乳腺癌, 增殖, 迁移, 侵袭

Abstract: Background and purpose: MicroRNA (miRNA) is closely related to the occurrence and development of tumors. This study aimed to investigate the expression of miR-6775-3p in breast cancer cells and its effect on the biological characteristics of breast cancer cells. Methods: Real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) was used to detect the expression level of miR-6775-3p in four breast cancer cell lines MDA-MB-231, MDA-MB-453, MDA-MB-468 and BT-549. After miR-6775-3p was over-expressed in breast cancer cells with the lowest expression level of miR-6775-3p, the proliferation of cells was detected by cell counting kit-8 (CCK-8) assay, and the migration and invasion ability of cells were detected by transwell migration and invasion assay. To further explore the molecular mechanism of miR-6775-3p in breast cancer, the expressions of cyclin-dependent protein kinase 4 (CDK4), CDK6 and invasion and metastasis markers matrix metalloproteinase 17 (MMP17) and MMP24 in miR-6775-3p overexpressing breast cancer cells were detected by RTFQ-PCR and Western blot. Results: RTFQ-PCR results showed that the expression of miR-6775-3p was the lowest in breast cancer cell line MDA-MB-453, and the expression level of miR-6775-3p was significantly increased after transfection of miR-6775-3p mimics in MDA-MB-453 cells (P<0.001). CCK-8 assay results showed that the cell proliferation activity was significantly decreased after miR-6775-3p was overexpressed in MDA-MB-453 cells (P<0.01). Transwell migration and invasion assay results showed that the cell migration (P<0.001) and invasion abilities (P<0.01) were significantly decreased after miR-6775-3p was over-expressed in MDA-MB-453 cells. RTFQ-PCR and Western blot results showed that the expressions of cyclin-dependent protein kinases CDK4 and CDK6, as well as the mRNA and protein levels of cell invasion and metastasis markers MMP17 and MMP24 were significantly decreased (P<0.01). Conclusion: miR-6775-3p may inhibit the proliferation, migration and invasion of breast cancer cells.

Key words: miR-6775-3p, Breast cancer, Proliferation, Migration, Invasion