南方医科大学学报

南方医科大学学报 ›› 2020, Vol. 40 ›› Issue (06): 869-875.doi: 10.12122/j.issn.1673-4254.2020.06.15

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MiR-204通过靶向调控HNRNPA2B1抑制乳腺癌的侵袭和转移

张丽萍，白 俊，胡雅琼，周丹丹，郑 荃，尹崇高，牟青杰，李洪利   

  • 出版日期:2020-06-20 发布日期:2020-06-20
  • 基金资助:

MiR-204 inhibits invasion and metastasis of breast cancer cells by targeted regulation of HNRNPA2B1

  

  • Online:2020-06-20 Published:2020-06-20

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摘要： 目的 探讨miR-204通过靶向调控HNRNPA2B1对乳腺癌侵袭和转移的影响。方法 生物信息学数据库查询miR-204在乳腺癌患者中的表达情况和 miR-204对乳腺癌患者生存率的影响；RT-qRCR检测miR-204在乳腺癌细胞系中的表达情况；利用 GV369-miR-204过表达慢病毒上调MDA-MB-231细胞中的miR-204；Transwell实验检测miR-204对MDA-MB-231细胞侵袭和迁移能力的影响；生物信息学方法确定miR-204的关键基因（Hub基因）；双荧光素酶实验检测miR-204和HNRNPA2B1的靶向关系；Western blot实验检测过表达miR-204后HNRNPA2B1的表达情况；Transwell实验检测MDA-MB-231/miR-204+NC、MDA-MB-231/miR-204+HNRNPA2B1细胞的侵袭和迁移能力的变化。结果 miR-204在乳腺癌组织中表达降低（P<0.05），低表达水平的miR-204与患者不良预后相关（P<0.05）；miR-204在乳腺癌细胞系MDA-MB-231细胞系中表达量低于正常乳腺上皮细胞MCF-10A的表达（P<0.0001）。通过过表达慢病毒上调MDA-MB-231细胞中miR-204的表达，结果显示上调miR-204可抑制乳腺癌细胞的侵袭和迁移能力（P<0.05）；HNRNPA2B1为miR-204的Hub基因，starbase网站发现miR-204-5p 和HNRNPA2B1表达呈负相关且HNRNPA2B1在乳腺癌患者中表达升高（P<0.05）；生存分析证明高表达HNRNPA2B1的乳腺癌患者预后不良（P<0.05）；双荧光素酶实验证明miR-204和HNRNPA2B1可靶向结合（P<0.05）。Western blot和Transwell实验证明，miR-204通过靶向调控HNRNPA2B1抑制乳腺癌的侵袭和迁移，且差异具有统计学意义（P<0.05）。结论 miR-204在乳腺癌组织和细胞中表达降低，且miR-204可通过靶向调控HNRNPA2B1抑制乳腺癌的侵袭和转移。

Abstract: Objective To investigate the effect of miR-204 on the invasion and metastasis of breast cancer by targeted regulation of HNRNPA2B1. Methods The bioinformatics database was used to obtain data of the expressions of miR-204 in breast cancer patients and the survival rate of the patients. RT-qPCR was used to detect the expression of miR-204 in breast cancer cell lines. The expression vector GV369-miR-204 was used to overexpress miR-204 in MDA-MB-231 cells. Transwell assay was performed to detect the effect of miR-204 on the migration and invasion ability of the breast cancer cells. The key genes (hub genes) of miR-204 were determined by bioinformatics method. A dual luciferase assay was used to analyze the targeting relationship between miR-204 and HNRNPA2B1. The expression of HNRNPA2B1 in MDA-MB-231 cells after miR-204 overexpression was detected by Western blotting, and Transwell assay was used to examine the changes in the cell invasion ability. Results The expression of miR-204 was decreased in both breast cancer tissues, and was significantly lower in breast cancer MDA-MB-231 cells than in MCF-10A cells (P<0.05). The decreased expression of miR-204 was associated with poorer prognosis of breast cancer patients (P<0.05). Upregulation of miR-204 in MDA-MB-231 cells significantly inhibited the invasion and migration of the cells (P<0.05). Analysis of the data from the Starbase revealed that the expression of miR-204-5p was negatively correlated with the expression of HNRNPA2B1, and the expression of HNRNPA2B1 was increased in breast cancer patients (P<0.05) in association with a poorer prognosis of the patients (P<0.05). Dual luciferase assay demonstrated that miR-204 could bind to HNRNPA2B1 in a target-specific manner. Western blotting and Transwell assay showed that miR-204 significant inhibited the migration and invasion ability of breast cancer cells by targeting HNRNPA2B1 (P<0.05). Conclusion miR-204 expression is decreased in breast cancer tissues and cells, and its overexpression can inhibit the invasion and metastasis of breast cancer cells by targeted regulation of HNRNPA2B1.