miR-196通过调控上皮-间充质转化抑制骨肉瘤细胞侵袭  

miR-196 inhibits the invasion of osteosarcoma cells by regulating epithelial-mesenchymal transition

作　　者：张靖勇 贺明[1] ZHANG Jingyong;HE Ming(Department of Orthopaedic Surgery,Shengjing Hospital,China Medical University,Liaoning Shenyang 110000,China)

机构地区：[1]中国医科大学附属盛京医院骨科,辽宁沈阳110000

出　　处：《现代肿瘤医学》2023年第22期4136-4141,共6页Journal of Modern Oncology

基　　金：辽宁省自然科学基金(编号:2020-MS-177);盛京医院345人才计划(40B)。

摘　　要：目的:探究miR-196在骨肉瘤和瘤旁组织中的表达差异,进而研究miR-196在骨肉瘤MG-63细胞中对SMAD4和上皮-间充质转化相关因子的调控作用及其对细胞侵袭能力的影响。方法:RT-qPCR检测20例患者的骨肉瘤和瘤旁组织中miR-196的差异性表达,Targetscan和miRDB数据库寻找miR-196序列与SMAD4序列是否存在结合位点,Spearman相关系数分析miR-196与SMAD4的表达相关性,双荧光素酶报告基因实验验证miR-196对SMAD4的调控作用,RT-qPCR和Western blotting分别检测miR-196与SMAD4对上皮-间充质转化相关因子在RNA水平和蛋白质水平表达的调控关系,Transwell实验检测miR-196对MG-63细胞侵袭能力的影响。结果:骨肉瘤组织中miR-196表达低于瘤旁组织,miR-196和SMAD4在骨肉瘤中的表达呈负相关(Y=-0.241 8*X+1.188),过表达miR-196可以在RNA水平和蛋白质水平抑制SMAD4、N-cadherin和Vimentin的表达并促进E-cadherin的表达,同时还能抑制骨肉瘤MG-63细胞的侵袭能力。结论:miR-196通过下调SMAD4抑制上皮-间充质转化过程,发挥抑制骨肉瘤MG-63细胞侵袭能力的作用。Objective:To explore the differential expression of miR-196 in osteosarcoma and adjacent tissues,and research into the role of miR-196 on SMAD4 and epithelial-mesenchymal transformation related factors in osteosarcoma MG-63 cells and the effect of miR-196 on the invasion ability of osteosarcoma MG-63 cells.Methods:RT-qPCR was used to detect the differential expression of miR-196 in osteosarcoma and adjacent tissues of 20 patients.The Targetscan and miRDB database was used to search for the binding site of miR-196 sequence and SMAD4 sequence.Spearman correlation analysis was used to analyze the correlation between miR-196 and SMAD4 expression.The regulation of miR-196 and SMAD4 was verified by dual luciferase reporting assay.The expression relationships of miR-196 and SMAD4 and epithelial-mesenchymal transformation related factors at RNA level and protein level were detected by RT-qPCR and Western blotting respectively.Transwell assay was used to detect the effect of miR-196 on the invasion ability of osteosarcoma MG-63 cells.Results:miR-196 was down-regulated in osteosarcoma tissues compared with adjacent tissues,and the expression of miR-196 and SMAD4 was negatively correlated in osteosarcoma(Y=-0.241 8*X+1.188).Overexpression of miR-196 can inhibit the expressions of SMAD4,N-cadherin and Vimentin at both RNA and protein levels,it can also promote the expression of E-cadherin,and inhibit the invasion ability of osteosarcoma MG-63 cells.Conclusion:miR-196 can inhibit the progression of epithelial-mesenchymal transformation by down-regulating SMAD4,thus inhibiting the invasion ability of osteosarcoma MG-63 cells.

关 键 词：骨肉瘤 MICRORNAS 上皮-间充质转化 侵袭 

分 类 号：R738.1[医药卫生—肿瘤]