| 陈夫圆,孙利平,朱远见,等.微小 RNA-490-3p靶向 FK506相关蛋白 14表达抑制骨肉瘤 MG63细胞侵袭和迁移的实验研究[J].安徽医药,2022,26(10):2063-2067. |
| 微小 RNA-490-3p靶向 FK506相关蛋白 14表达抑制骨肉瘤 MG63细胞侵袭和迁移的实验研究 |
| Inhibitory effect of miR-490-3p targeting FKBP14 on migration and invasion of osteosarcoma MG63 cells |
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| DOI：10.3969/j.issn.1009-6469.2022.10.036 |
| 中文关键词: 骨肉瘤 骨肉瘤 MG63 微小 RNA-490-3p 细胞侵袭 迁移 FK506相关蛋白 14 |
| 英文关键词: Osteosarcoma Osteosarcoma MG63 miR-490-3p Invasion Migration FKBP14 |
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| 中文摘要: |
| 目的探讨微小 RNA-490-3p(miR-490-3p)对骨肉瘤 MG63细胞侵袭、迁移的影响及其机制。方法 2019年 9月至 2020年 4月，从中科院上海细胞库购买人骨肉瘤细胞株 MG63进行体外培养，分为对照组(未转染)miR-NC组(转染 miR-NC)、 miR-490-3p组(转染 miR-490-3p mimics)，miR-490-3p+pcDNA组(共转染 miR-490-3p mimics与空载体、)和 miR-490-3p+FKBP14组(共转染 miR-490-3p mimics与 FKBP14过表达载体)，采用 RT-PCR检测 miR-490-3p表达， Transwell小室法检测细胞侵袭、迁移，蛋白质印迹法检测钙黏蛋白 E(E-cadherin)、基质金属蛋白酶 -2(MMP-2)、上皮间质转化因子 Twist转录因子( Twist)和 FK506相关蛋白 14(FKBP14)蛋白表达，双荧光素酶报告基因实验检测 miR-490-3p和 FKBP14的靶向关系。结果与对照组比较， miR-490-3p组细胞中 miR-490-3p表达水平明显升高( 3.68±0.37比 1.02±0.10，P<0.05)，而侵袭细胞数( 33.25±2.02比 |
| 英文摘要: |
| Objective To investigate the effect of miR-490-3p on the invasion and migration of osteosarcoma MG63 cells and its mechanism.Methods From September 2019 to April 2020,human osteosarcoma cell line MG63 was purchased from Shanghai cellbank, Chinese Academy of Sciences, cultured in vitro and assigned into control group (untransfected), miR-NC group (transfected with miR-NC), miR-490-3p group (transfected with miR-490-3p mimics), miR-490-3p+pcDNA group (co-transfected miR-490-3p mimics and empty vector) and miR-490-3p+FKBP14 group (co-transfected miR-490-3p mimics and FKBP14 overexpression vector). The ex. pression of miR-490-3p was detected by RT-PCR; the invasion and migration of MG63 cells were detected by Transwell; the protein ex. pressions of E-cadherin, Matrix metalloproteinase-2 (MMP-2), epithelial-mesenchymal transforming factor Twist transcription factor (Twist) and FK506-associated protein 14 (FKBP14) were detected by Western blotting. The targeting relationship of miR-490-3p and FKBP14 was determined by double luciferase reporter gene assay.Results Compared with the control group, the expression level of miR-490-3p in the cells of the miR-490-3p group was significantly increased [(3.68±0.37) vs. (1.02±0.10), P<0.05], while the number of invaded cells [(33.25±2.02) vs. (76.52±6.00), P<0.05], number of migrating cells [(45.56±3.38) vs. (97.85±6.45), P<0.05] and the pro. tein expression levels of MMP-2, Twist and FKBP14 reduced, and the protein expression level of E-cadherin increased, while there was no significant difference between the miR-NC group and the control group (P>0.05). The results of double luciferase reporter gene ex. periment confirmed that FKBP14 was a potential target gene of miR-490-3p. Compared with the miR-490-3p+pcDNA group, the num. ber of invaded cells, number of migrating cells and the protein expression levels of MMP-2, Twist and FKBP14 in the miR-490-3p + FKBP14 group increased, while the protein expression level of E-cadherin reduced (P<0.05).Conclusion miR-490-3p could inhibittargeted FKBP14 expression to reduce the invasion and migration of osteosarcoma MG63 cells, which can be used as a molecular targetfor osteosarcoma treatment. |
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