论著 Original Article

miR-580对乳腺癌细胞系中 Twist1的调节作用

Published at: 2015年第35卷第6期

孟庆威 1 , 孙嘉 1 , 纪春连 1 , 席玉慧 2
1 哈尔滨医科大学附属肿瘤医院内四科,哈尔滨 150081
2 哈尔滨医科大学病理生理教研室,哈尔滨 150081
通讯作者 庆威 孟 Email: mqwei@ems.hrbmu.edu.cn
DOI: 10.3978/j.issn.2095-6959.10.3978/j.issn.2095-6959.2015.06.038
基金:
黑龙江省教育厅面上项目 12521236

摘要

目的:探讨miR-580在MCF-10A细胞系中对Twist1的调节。方法:本实验利用生物信息学方法预测Twist1的靶miRNA是miR-580。首先,采用qPCR法检测在MCF-10A系列细胞系中Twist1及miR-580的表达。然后,在MCF-10A细胞系中分别转染miR-580类似物和miR-580抑制物后,利用RT-PCR、Western blot、t检验分析Twist1的表达及细胞迁移能力的变化。最后利用荧光素酶实验验证miR-580通过结合在Twist1的3'UTR调节其表达。结果:1)在MCF-10A细胞系中Twist1与miR-580的表达呈负相关;2)在MCF-10A细胞系中转染miR-580类似物后,Twist1的表达下调;在MCF-10A细胞系中转染miR-580抑制物后Twist1的表达上调;3)在MCF-10A细胞系中引入miR-580类似物后细胞迁移能力降低;4)miR-580直接结合在Twist1的3'UTR。结论:miRNA-580在MCF-10A细胞系中通过结合在Twist1的3'UTR负向调节Twist1的表达从而克制细胞的迁移。


Twist1 regulation by miR-580 in breast cancer cell lines

Abstract

Objective: To explore the role of miR-580 in regulating Twist1 expression in MCF-10A cell line. Methods: By bioinformatics, we postulated that miR-580 is one of the regulators of Twist1. First, qPCR was used to evaluate the expression of Twist1 and miR-580 in MCF-10A series cell lines. Then, we transfected miR-580 analogs and inhibitors into the MCF-10A cells, respectively, the expression of Twist1 and the cell migration were detected by using RT-PCR, western blot and student’s t-test. Finally, by using luciferase experiments to verify the miR-580 can regulate the expression of Twist1 by binding to its 3'UTR. Results: 1) The expression of Twist1 is negatively correlated with miR-580 in MCF-10A cell lines; 2) transfected miR-580 analogues into MCF-10A cells can down-regulate the expression of Twist1, whereas transfected with miR-580 inhibitor can upregulate the expression of Twist1; 3) introduction of miR-580 analogues reduces cell migration in MCF-10A cell line; 4) miR-580 directly binds to Twist1 3'UTR. Conclusion: miRNA-580 regulates the expression of Twist1 and migration by binding to its 3'UTR in MCF-10A cell line.


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引用

引用本文: 庆威 孟, 嘉 孙, 春连 纪, 玉慧 席. miR-580对乳腺癌细胞系中 Twist1的调节作用[J]. 临床与病理杂志, 2015, 35(6): 1065-1069.
Cite this article as: MENG Qingwei, SUN Jia, JI Chunlian, XI Yuhui . Twist1 regulation by miR-580 in breast cancer cell lines[J]. Journal of Clinical and Pathological Research, 2015, 35(6): 1065-1069.