miRNA-192-5p在多发性骨髓瘤中的表达水平及其调控机制研究

Expression of miRNA-192-5p in multiple myeloma and its regulatory mechanism

目的探讨miR-192-5p在多发性骨髓瘤中的表达水平及其调控机制。方法选取骨髓瘤细胞株U266作为研究对象,利用荧光定量PCR检测多发性骨髓瘤细胞中miR-192-5p的表达水平;使用生物信息学网站预测miR-192-5p潜在靶基因USP1并利用双荧光素酶报告实验验证其靶向关系;利用细胞转染实验对多发性骨髓瘤细胞株进行miR-192-5p mimics、miR-192-5p inhibitor及相关对照组的细胞转染,采用CCK-8法和流式细胞仪分别检测转染后细胞的增殖和凋亡情况;利用Western blot检测转染后细胞中USP1蛋白表达水平。结果qRT-PCR结果显示,转染后多发性骨髓瘤细胞中miR-192-5p表达水平低于正常骨髓细胞(P<0.01);CCK-8细胞增殖实验、细胞划痕实验、Transwell体外侵袭转移实验和流式细胞仪检测实验结果显示,过表达miR-192-5p抑制骨髓瘤细胞的增殖、迁移、侵袭,促进骨髓瘤细胞的凋亡;靶基因预测及验证实验结果显示USP1是miR-192-5p的靶基因,miR-192-5p的表达可以显著降低骨髓瘤细胞中USP1蛋白的表达。结论miR-192-5p在多发性骨髓瘤中可能作为抑癌基因发挥作用,通过靶向作用于USP1抑制多发性骨髓瘤的发展。

Objective To explore the expression of miR-192-5p in multiple myeloma and its regulatory mechanism.Methods The Myeloma cell line U266 was selected as the object of study,and the expression of miR-192-5p in multiple myeloma cells was detected by fluorescence quantitative PCR.The potential target gene USP1 of miR-192-5p was predicted by bioinformatics website and its targeting relationship was verified by double luciferase reporting experiment.Multiple myeloma cell lines were transfected with miR-192-5p mimics,miR-192-5p inhibitor and associated control groups by cell transfection experiment.The proliferation and apoptosis of the transfected cells were detected by CCK-8 and flow cytometry respectively.The expression of USP1 protein in the transfected cells was detected by Western blot.Results The expression of miR-192-5p in the transfected multiple myeloma cells was lower than that in the normal bone marrow cells(P<0.01).The over-expression of miR-192-5p inhibited the proliferation,migration and invasion of myeloma cells and promoted the apoptosis of myeloma cells.The results of target gene prediction and validation experiments showed that USP1 was the target gene of,and the miR-192-5p could significantly reduce the expression of USP1 protein in myeloma cells.Conclusion miR-192-5p may play a role as a tumor suppressor gene in multiple myeloma and inhibit the development of multiple myeloma by targeting USP1.