| 韦杰合,韦仁杰,周业修.微小 218?5p调控 Wnt家族成员 2B抑制骨肉瘤细胞 143B的增殖、迁移和侵袭[J].安徽医药,2021,25(1):13-16. |
| 微小 218?5p调控 Wnt家族成员 2B抑制骨肉瘤细胞 143B的增殖、迁移和侵袭 |
| MiR?218?5p inhibits the proliferation，migration and invasion of osteosarcoma cells 143B via regulating Wnt family member 2B |
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| DOI：10.3969/j.issn.1009?6469.2021.01.004. |
| 中文关键词: 骨肉瘤 微小 RNA?218?5p Wnt蛋白质类 s 增殖 侵袭 P＜0.05。应用TargetSca，测，，结果显示Wnt家族，)， |
| 英文关键词: Osteosarcoma miR?218?5p Wnt Proteins Proliferation Invasion |
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| 中文摘要: |
| 目的探讨微小 RNA?218?5p(miR?218?5p)对骨肉瘤细胞增殖，侵袭和迁移能力的影响及其作用机制。方法 2016年 1月至 2019年 1月，实时荧光定量 PCR检测 miR?218?5p在骨肉瘤与正常骨组织及正常成骨细胞株与骨肉瘤细胞株的表达差异。应用 CCK?8法检测过表达 miR?218?5p对人骨肉瘤细胞株 143B增殖的影响；细胞划痕实验检测和 Transwell小室实验检测过表达 miR?218?5p对骨肉瘤细胞 143B迁移能力的影响。同时，应用在线 miRNA靶基因预测软件预测 miR?218?5p的靶基因，并应用荧光素酶报告实验进行结合位点验证。蛋白质印迹法验证靶基因的蛋白表达水平。结果 miR?218?5p在骨肉瘤组织中的表达水平(0.59±0.21)%明显低于正常骨组织，骨肉瘤细胞系中的表达水平(0.712±0.015)%、(0.685±0.016)%、(0.542±0.021)%、(0.524±0.011)%明显低于正常成骨细胞， P＜0.05。同时，与对照组相比，过表达 miR?218?5p(miR?218?5p mimic)的骨肉瘤细胞 143B的细胞活力 t6h(0.274±0.011)t24h(0.514±0.04)t48h(0.828±0.16)t72h(1.212±0.013)侵袭(164±12)个和迁移距离(26±2.4)%明显减弱， n网站进行预成员 2B(WNT2B为 miR?218?5p的潜在靶基因之一；荧光素酶报告实验结果表明 miR?218?5p直接靶向 WNT2B；蛋白质印迹法证实 miR?218?5p过表达组细胞中 WNT2B蛋白的表达水平较对照组明显减少。结论 miR?218?5p在人骨肉瘤组织和癌细胞中低表达，其可能通过靶向 WNT2B抑制骨肉瘤细胞 143B的增殖、迁移和侵袭。 |
| 英文摘要: |
| Objective To investigate the effect of miR?218?5p on proliferation，invasion and migration of osteosarcoma cells and its mechanism.Methods From January 2016 to January 2019，the expression differences of miR?218?5p in osteosarcoma and nor?mal bone tissue and normal osteoblasts cell lines and osteosarcoma cell lines were detected by real?time fluorescence quantitativePCR.While the effect of miR?218?5p overexpression in osteosarcoma cells 143B were investigated using CCK?8.The effects of miR?218?5p overexpression on osteosarcoma cells 143B migration and invasion was tested using wound healing assay and transwell as?say.Meanwhile，the target genes of miR?218?5p were predicted by online miRNA target gene prediction software，and the binding sites were verified by luciferase reporting experiment.Results The expression level of miR?218?5p in osteosarcoma tissues(0.59±0.21)% was significantly lower than that in adjacent tissues，and the expression level in osteosarcoma cell lines(0.712±0.015)%，(0.685±0.016)%，(0.542±0.021)%，(0.524±0.011)% was significantly lower than that in normal osteoblast cells(P＜0.05).Mean? while，compared with the control group，143B cells of osteosarcoma cells overexpressing miR?218?5p mimic showed significantly re? duced cell activity t6h(0.274±0.011)，t24h(0.514±0.04)，t48h(0.828±0.16)，t72h(1.212±0.013)，invasion(164±12)，and migration dis? tance(26±2.4)%，P＜0.05.TargetScan was used for prediction，and the results showed that Wnt family member 2B(WNT2B)was one of the potential target genes of miR?218?5p.Luciferase reporting results showed that miR?218?5p directly targeted WNT2B.West?ern blot confirmed that the expression level of WNT2B protein in the miR ?218?5P overexpression group was significantly lowerthan that in the control group.Conclusion miR?218?5p is low expressed in human osteosarcoma tissues and cancer cells，miR?218? 5p plays as suppressor in osteosarcoma，which inhibiting proliferation，migration and invasion of osteosarcoma cells 143B via target? ing WNT2B. |
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