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  • 于金玲,高蓓敏,沈卫达,等.Hsa_circ_USP42竞争性结合miR-182-5p抑制三阴性乳腺癌转移的机制研究[J].同济大学学报(医学版),2020,41(2):205-210.    [点击复制]
  • YU Jin-ling,GAO Bei-min,SHEN Wei-da,et al.Hsa_circ_USP42 inhibits metastasis in triple negative breast cancer through competitive binding miR-182-5p[J].同济大学学报(医学版),2020,41(2):205-210.   [点击复制]
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Hsa_circ_USP42竞争性结合miR-182-5p抑制三阴性乳腺癌转移的机制研究
于金玲,高蓓敏,沈卫达,崔静,王浩峰,朱江帆
0
(同济大学附属东方医院普外科,上海200120; 上海市长宁区妇幼保健院乳腺科,上海200051)
摘要:
目的探索hsa_circ_USP42作为miR-182-5p海绵在TNBC中的生物学功能及调控机制。方法RT-PCR技术检测hsa_circ_USP42在TNBC癌组织及癌旁组织中表达;通过双荧光素酶报告基因分析等技术研究hsa_circ_USP42和miR-182-5p的相互作用;过表达hsa_circ_USP42,qPCR技术及Western印迹法检测靶miRNA miR-182-5p及靶基因MTSS1的表达水平;Transwell及CCK8实验检测hsa_circ_USP42对细胞增殖、侵袭和迁移的影响。结果Hsa_circ_USP42在TNBC中表达明显下调(P=0.000),淋巴结阳性组表达低于淋巴结阴性组(P=0.003)。双荧光素酶实验证实hsa_circ_USP42与靶miR-182-5P可有效结合;细胞功能实验表明,过表达hsa_circ_USP42可抑制TNBC细胞迁移、侵袭和增殖。体外实验表明,过表达hsa_circ_USP42后,靶miRNA miR-182-5p上调,而靶基因MTSS1在转录水平及蛋白水平均降低。结论Hsa_circ_USP42在TNBC中低表达,与miR-182-5p有效结合,可发挥miRNA海绵上调MTSS1表达,抑制TNBC侵袭及转移,是参与TNBC生物学行为机制之一。
关键词:  三阴性乳腺癌  hsa_circ_USP42  miR-182-5p  转移
DOI:10.16118/j.1008-0392.2020.02.011
投稿时间:2019-12-02
基金项目:上海市长宁区科学技术委员会课题(CNKW20164Y13)
Hsa_circ_USP42 inhibits metastasis in triple negative breast cancer through competitive binding miR-182-5p
YU Jin-ling,GAO Bei-min,SHEN Wei-da,CUI Jing,WANG Hao-feng,ZHU Jiang-fa
(Dept. of General Surgery, East Hospital, Tongji University School of Medicine, Shanghai 200120, China;Dept. of Breast Surgery, Shanghai Changning Maternity & Infant Health Hospital, Shanghai 200051, China)
Abstract:
ObjectiveTo investigate the biological function of hsa_circ_USP42 as sponge of miR-182-5p in triple negative breast cancer(TNBC). MethodsThe expression of hsa_circ_USP42 was detected by qRT-PCR in cancer tissue samples from 30 TNBC patients. Cell migration assay was conducted using Transwell assay, and cell proliferation was evaluated with the Cell Counting Kit 8(CCK8)in breast cancer MDA-MB-231 cells. Bioinformatics prediction was performed to find the candidate hsa_circ_USP42 that may target miR-182-5p. Luciferase assays using miR-182 5'-untranslated region(5'-UTR)reporters were conducted to confirm the binding of miR-182-5p to the hsa_circ_USP42. Real-time quantitative RT-PCR(qPCR)and Western blotting were used to detect the expression of miR-182-5p and MTSS1 in MDA-MB-231 cells affected by over expression of hsa_circ_USP42. ResultsRT-PCR showed that the expression of hsa_circ_USP42 in TNBC was significantly lower than that in adjacent tissues(P=0.000). In vitro experiments showed that overexpression of hsa_circ_USP42 significantly enhanced the proliferation and migration of MDA-MB-231 cells. RT-PCR and Western blotting analysis showed that overexpression of hsa_circ_USP42 enhanced MTSS1 expression at both mRNA and protein levels, and reduced the expression of miR-182-5p. ConclusionThe over-expression of hsa_circ_USP42 in TNBC can inhibit cell proliferation and migration, which may be serve as an sponge of miR-182 to enhance expression of MTSS1 that inhibits metastasis of TNBC.
Key words:  triple-negative breast cancer  hsa_circ_USP42  microRNA-182  migration

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