miR-580对乳腺癌细胞系中 Twist1的调节作用
| 作者: |
1孟庆威,
1孙嘉,
1纪春连,
2席玉慧
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: |
孟庆威
Email: mqwei@ems.hrbmu.edu.cn |
| DOI: | 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。
关键词：
乳腺癌
Twist1 蛋白
miR-580
细胞迁移
Twist1 regulation by miR-580 in breast cancer cell lines
CorrespondingAuthor: MENG Qingwei Email: mqwei@ems.hrbmu.edu.cn
DOI: 10.3978/j.issn.2095-6959.2015.06.038
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.