临床与病理杂志
臨床與病理雜誌
림상여병리잡지
International Journal of Pathology and Clinical Medicine
2015年
6期
1065-1069
,共5页
孟庆威%孙嘉%纪春连%席玉慧
孟慶威%孫嘉%紀春連%席玉慧
맹경위%손가%기춘련%석옥혜
乳腺癌%Twist1蛋白%miR-580%细胞迁移
乳腺癌%Twist1蛋白%miR-580%細胞遷移
유선암%Twist1단백%miR-580%세포천이
Breast cancer%Twist1%miR-580%cell migration
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。
目的：探討miR-580在MCF-10A細胞繫中對Twist1的調節。方法：本實驗利用生物信息學方法預測Twist1的靶miRNA是miR-580。首先，採用qPCR法檢測在MCF-10A繫列細胞繫中Twist1及miR-580的錶達。然後，在MCF-10A細胞繫中分彆轉染miR-580類似物和miR-580抑製物後，利用RT-PCR、Western blot、t檢驗分析Twist1的錶達及細胞遷移能力的變化。最後利用熒光素酶實驗驗證miR-580通過結閤在Twist1的3'UTR調節其錶達。結果：1)在MCF-10A細胞繫中Twist1與miR-580的錶達呈負相關；2)在MCF-10A細胞繫中轉染miR-580類似物後，Twist1的錶達下調；在MCF-10A細胞繫中轉染miR-580抑製物後Twist1的錶達上調；3)在MCF-10A細胞繫中引入miR-580類似物後細胞遷移能力降低；4)miR-580直接結閤在Twist1的3'UTR。結論：miRNA-580在MCF-10A細胞繫中通過結閤在Twist1的3'UTR負嚮調節Twist1的錶達從而剋製細胞的遷移。
목적：탐토miR-580재MCF-10A세포계중대Twist1적조절。방법：본실험이용생물신식학방법예측Twist1적파miRNA시miR-580。수선，채용qPCR법검측재MCF-10A계렬세포계중Twist1급miR-580적표체。연후，재MCF-10A세포계중분별전염miR-580유사물화miR-580억제물후，이용RT-PCR、Western blot、t검험분석Twist1적표체급세포천이능력적변화。최후이용형광소매실험험증miR-580통과결합재Twist1적3'UTR조절기표체。결과：1)재MCF-10A세포계중Twist1여miR-580적표체정부상관；2)재MCF-10A세포계중전염miR-580유사물후，Twist1적표체하조；재MCF-10A세포계중전염miR-580억제물후Twist1적표체상조；3)재MCF-10A세포계중인입miR-580유사물후세포천이능력강저；4)miR-580직접결합재Twist1적3'UTR。결론：miRNA-580재MCF-10A세포계중통과결합재Twist1적3'UTR부향조절Twist1적표체종이극제세포적천이。
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. hTen, 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’st-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) hTe 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.
