miR-147a靶向MCM3抑制脉络膜黑色素瘤细胞增殖、迁移和侵袭  

Effects of miR-147a targeting MCM3 on proliferation,migration and invasion of choroidal melanoma cells

作　　者：刘然 应曼曼 张晓楠 宁宏[1] LIU Ran;YING Manman;ZHANG Xiaonan;NING Hong(Department of Ophthalmology,The First Hospital of China Medical University,Liaoning Shenyang 110001,China.)

机构地区：[1]中国医科大学附属第一医院眼科,辽宁沈阳110001

出　　处：《现代肿瘤医学》2021年第12期2024-2029,共6页Journal of Modern Oncology

基　　金：辽宁省自然科学基金项目(编号:20180551171)。

摘　　要：目的:研究miR-147a对脉络膜黑色素瘤细胞增殖、迁移和侵袭的影响,并探究其作用机制。方法:采用脂质体转染人脉络膜黑色素瘤MUM-2B细胞,设置miR-NC、miR-147a mimics、inhibitor-NC、miR-147a inhibitor四组。采用EDU染色法检测细胞增殖;Transwell实验检测细胞迁移、侵袭;Targetscan生物信息学网站预测分析miR-147a下游靶基因;双荧光素酶报告基因实验验证miR-147a是否靶向结合MCM3 mRNA 3'UTR区;蛋白质印迹法(Western Blot)检测靶基因MCM3蛋白表达。结果:过表达miR-147a可显著降低MUM-2B细胞的增殖数、迁移数和侵袭数,相反,敲减miR-147a提高细胞增殖数、迁移数和侵袭数。miR-147a直接靶向并负向调控MCM3蛋白表达。沉默MCM3可抑制MUM-2B细胞增殖、迁移和侵袭。结论:miR-147a通过直接靶向MCM3抑制脉络膜黑色素瘤MUM-2B细胞的增殖、迁移和侵袭。miR-147a/MCM3轴可能是治疗脉络膜黑色素瘤的新靶标。Objective:To study the effects of miR-147a on the proliferation,migration and invasion of choroidal melanoma cells,and to explore its mechanism.Methods:The human choroidal melanoma MUM-2B cells were transfected by liposome transfection and were divided into miR-NC group,miR-147a mimics group,inhibitor-NC group and miR-147a inhibitor group.The ability of cell proliferation was detected by EDU staining.The ability of cell migration and invasion was detected by Transwell test.The downstream target genes of miR-147a were predicted and analyzed by Targetscan bioinformatics website.The targeting relationship between miR-147a and MCM3 mRNA 3'UTR region was verified by dual luciferase reporter gene assay.The protein expression of target gene MCM3 was detected by Western Blot.Results:Overexpression of miR-147a significantly decreased the numbers of proliferation,migration and invasion of MUM-2B cells.On the contrary,suppression of miR-147a increased the numbers of proliferation,migration and invasion of cells.miR-147a directly targeted and regulated the expression of MCM3 protein.Overexpression of miR-147a inhibited MCM3 protein levels,and knocking down miR-147a promoted MCM3 levels.Knocking down MCM3 can inhibit the proliferation,migration and invasion of MUM-2B cells.Conclusion:miR-147a inhibits the proliferation,migration and invasion of choroidal melanoma MUM-2B cells by directly targeting MCM3.The miR-147a/MCM3 axis may be a new target for the treatment of choroidal melanoma.

关 键 词：miR-147a MCM3 脉络膜黑色素瘤 细胞增殖 迁移 侵袭 

分 类 号：R739.7[医药卫生—肿瘤]