中国癌症杂志 ›› 2020, Vol. 30 ›› Issue (9): 667-673.doi: 10.19401/j.cnki.1007-3639.2020.09.005

• 论著 • 上一篇    下一篇

miR-26b-3p对乳腺癌细胞生物学行为的影响及作用机制研究

田连芳,李丽玮,张小冲,郑丽春,左江华,孙春秀,刘登湘   

  1. 邢台市人民医院检验科,河北 邢台 054001
  • 出版日期:2020-09-30 发布日期:2020-10-10
  • 通信作者: 刘登湘 E-mail: rmyy666@163.com
  • 基金资助:
    邢台市科技计划项目(2012ZC085)。

Effect of miR-26b-3p on biological behaviors of breast cancer cells and its molecular mechanism

TIAN Lianfang, LI Liwei, ZHANG Xiaochong, ZHENG Lichun, ZUO Jianghua, SUN Chunxiu, LIU Dengxiang   

  1. Department of Clinical Laboratory, Xingtai People’s Hospital, Xingtai 054001, Hebei Province, China
  • Online:2020-09-30 Published:2020-10-10
  • Contact: LIU Dengxiang E-mail: rmyy666@163.com

摘要: 背景与目的:miRNA是一类长度为21~23个核苷酸的单链非编码RNA分子,其作用机制主要为靶向于mRNA的3’非翻译区(3’ untranslated region,3’UTR)从而抑制其靶基因的表达。miRNA在肿瘤的发生、发展过程中发挥着关键作用,探讨miR-26b-3p对乳腺癌生物学行为的影响及作用机制。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miR-26b-3p在三种乳腺癌细胞系MCF-7、MDA-MB-231和MDA-MB-453中的表达,选取miR-26b-3p表达水平最低的乳腺癌细胞转染miR-26b-3p mimics后,采用细胞计数试剂盒(cell counting kit-8,CCK-8)法检测细胞的增殖,采用transwell迁移和侵袭实验检测细胞迁移和侵袭能力,通过小动物活体成像及裸鼠移植瘤模型检测miR-26b-3p对乳腺癌细胞裸鼠移植瘤生长和转移的影响,采用双荧光素酶报告基因分析检测miR-26b-3p与锌指E盒结合同源盒基因1(zinc finger E-box binding homeobox 1,ZEB1)的相互作用,采用RTFQ-PCR和蛋白质印迹法(Western blot)检测ZEB1的表达。结果:乳腺癌细胞系MDA-MB-453中miR-26b-3p表达最低,在MDA-MB-453细胞中转染miR-26b-3p mimics后,miR-26b-3p的表达水平显著升高(P<0.05),细胞的增殖能力显著降低(P<0.05),细胞的迁移(P<0.001)和侵袭能力(P<0.01)显著降低。过表达miR-26b-3p可抑制裸鼠体内乳腺癌移植瘤的生长和转移。miR-26b-3p可与ZEB1的3’UTR结合,抑制ZEB1的表达。结论:miR-26b-3p可靶向于ZEB1,抑制乳腺癌细胞的增殖、迁移和侵袭,抑制乳腺癌的生长和转移。

关键词: 乳腺癌, miR-26b-3p, 增殖, 迁移, 侵袭

Abstract: Background and purpose: miRNA is a type of single-stranded non-coding RNA molecule with a length of about 21-23 nucleotides, which mainly functions by targeting the 3’ untranslated region (3’UTR) of mRNA so as to inhibit the expression of its target gene. miRNA plays a key role in the occurrence and development of cancer. This study aimed to explore the effect of miR-26b-3p on the biological characteristics of breast cancer and its mechanism. Methods: Real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) was used to detect the expression level of miR-26b-3p in MCF-7, MDA-MB-231 and MDA- MB-453 breast cancer cell lines. miR-26b-3p mimics were transfected in the breast cancer cells with the lowest expression of miR-26b-3p. Cell proliferation was detected by cell counting kit-8 (CCK-8) assay, and cell migration and invasion abilities were detected by transwell migration and invasion assay. The effects of miR-26b-3p on the growth and metastasis of breast cancer xenografts in nude mice were detected by in vivo imaging system and in vivo metastasis model. Dual luciferase reporter assay was used to detect the interaction between miR-26b-3p and zinc finger E-box binding homeobox 1 (ZEB1). RTFQ-PCR and Western blot were used to detect the expression of ZEB1. Results: The expression of miR-26b-3p was the lowest in MDA-MB-453 cells. After transfection with miR-26b-3p mimics, the expression level of miR-26b-3p was significantly increased (P<0.05). After miR-26b-3p was overexpressed in MDA-MB-453 cells, the cell proliferation (P<0.05), migration (P<0.001) and invasion abilities (P<0.01) were significantly decreased. Overexpression of miR-26b-3p inhibited the growth and metastasis of breast cancer xenografts in nude mice. miR-26b-3p could bind the 3’UTR of ZEB1, and inhibit the expression of ZEB1. Conclusion: miR-26b-3p directly targets ZEB1 and inhibits the proliferation, migration and invasion of breast cancer cells. miR-26b-3p inhibits breast cancer growth and metastasis.

Key words: Breast cancer, miR-26b-3p, Proliferation, Migration, Invasion