摘要
目的:明确miR-214与MCL-1之间的靶向调控关系及其对骨髓瘤细胞侵袭迁移能力的影响。方法:qRT-PCR和Western blot检测miR-214和MCL-1在骨髓瘤细胞中的表达情况并做相关性分析;Transwell小室法检测上调miR-214或沉默MCL-1对骨髓瘤细胞株H929和U266迁移侵袭能力的影响;荧光素酶报告基因实验验证miR-214与MCL-1的靶向关系;Western blot检测上调或下调miR-214对MCL-1表达的影响。结果:miR-214在骨髓瘤细胞中表达下调,MCL-1则显上调,二者呈负相关;上调miR-214或敲低MCL-1可显著抑制骨髓瘤细胞H929和U266的侵袭迁移能力;荧光素酶报告基因实验证实MCL-1是miR-214的靶基因,Westerm blot结果发现miR-214负调控MCL-1表达;MCL-1的额外补充可逆转miR-214对骨髓瘤细胞H929和U266迁移侵袭能力的抑制作用。结论:miR-214通过负调控MCL-1表达抑制骨髓瘤细胞的迁移侵袭。
Objective:To clarify the relationship between microRNAs-214 and MCL-1 and their effects on the invasion and migration of myeloma cells.Methods:qRT-PCR and Western blot were used to detect the expression of miR-214 and MCL-1 in myeloma cells and analyze the correlation.Transwell cell assay was used to detect the effects of overexpression of miR-214 or silencing of MCL-1 on migration and invasion of myeloma cell lines H929 and U266.Luciferase reporter gene assay confirmed the target relationship between miR-214 and MCL-1.Western blot detected the effect of up regulation or down regulation of miR-214 on MCL-1 expression.Results:The expression of miR-214 was downregulated in myeloma cells,while MCL-1 was upregulated,and the two were negatively correlated.Up regulation of miR-214 or knockdown of MCL-1 significantly inhibited the invasion and migration of H929 and U266 in myeloma cells.Luciferase reporter gene assay confirmed that MCL-1 was the target gene of microRNAs-214.Western blot showed that miR-214 negatively regulated the expression of MCL-1.Additional supplementation of MCL-1 reversed the inhibitory effect of miR-214 on migration and invasion of myeloma cells H929 and U266.Conclusion:miR-214 inhibits migration and invasion of myeloma cells through negative regulation of MCL-1 expression.
作者
孙飞
王硕
赵倩
Sun Fei;Wang Shuo;Zhao Qian(Department of Orthopaedics,Zaozhuang Mining Group Zaozhuang Hospital,Shandong Zaozhuang 277100,China)
出处
《现代肿瘤医学》
CAS
2020年第5期740-746,共7页
Journal of Modern Oncology
