陈 静,董善武,陈咏丽,等.miR-640通过抑制SLIT1介导Wnt/β-catenin通路促进脑胶质瘤对替莫唑胺的耐药[J].肿瘤学杂志,2022,28(7):568-573.
miR-640通过抑制SLIT1介导Wnt/β-catenin通路促进脑胶质瘤对替莫唑胺的耐药
miR-640 Promote Proliferation, Invasion and Resistance to Temozolomide in Glioma by Inhibiting SLIT1-mediated Wnt/β-catenin Pathway
投稿时间:2021-12-29  
DOI:10.11735/j.issn.1671-170X.2022.07.B007
中文关键词:  脑胶质瘤  替莫唑胺  miR-640  Wnt/β-catenin通路
英文关键词:glioma  temozolomide  miR-640  Wnt/β-catenin pathway
基金项目:武汉市卫健委科研项目(WX21C05)
作者单位
陈 静 武汉市第四医院 
董善武 武汉市第四医院 
陈咏丽 武汉市第四医院 
陈海珊 武汉市第四医院 
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中文摘要:
      摘 要:[目的]评价miR-640介导Wnt/β-catenin通路对脑胶质瘤耐药细胞增殖、侵袭及替莫唑胺(temazolamide,TMZ)敏感性的影响。[方法] 利用实时荧光定量聚合酶链反应(RT-qPCR)测定脑胶质瘤细胞内miR-640的表达。U87/TMZ细胞分为NC inhibitor组、 miR-640 inhibitor组和miR-640 inhibitor+si-SLIT1组。用不同浓度(15、30、60、120、240 、480和960 μmol/L)TMZ处理细胞,检测细胞对TMZ的耐药性;运用细胞计数试剂盒8(CCK-8)、Transwell实验检测U87/TMZ细胞增殖活力和侵袭力;用双荧光素酶报告基因实验验证miR-640与SLIT1的靶向关系。采用Western blot检测Wnt/β-Catenin信号通路相关蛋白表达水平。[结果] miR-640在脑胶质瘤细胞中高表达,转染后miR-640 inhibitor组的miR-640表达水平、侵袭细胞数目(57±23 vs 153±31)、IC50值(131.61±8.97 vs 293.33±11.28)、增殖率(18.48±4.23 vs 43.84±8.94)均比NC inhibitor组低(P<0.05)。miR-640靶向调控SLIT1的表达,miR-640 inhibitor+si-SLIT1组β-catenin、c-myc、Cyclin D1蛋白表达水平均高于miR-640 inhibitor组(P<0.05)。[结论]miR-640通过SLIT1介导Wnt/β-catenin通路促进脑胶质瘤增殖、侵袭及对替莫唑胺的耐药性。
英文摘要:
      Abstract: [Objective] To investigate the effects of miR-640-mediated Wnt/β-catenin pathway on proliferation, invasion and sensitivity to temazolamide(TMZ) in glioma cells. [Methods] The expression of miR-640 in glioma cells as determined by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR). TMZ-resistant human glioma U87/TMZ cells were transfected with NC inhibitor, miR-640 inhibitor and miR-640 inhibitor+ si-SLIT1, respectively. U87/TMZ cells were treated with TMZ(15, 30, 60, 120, 240, 480 and 960 μmol/L) . Cell counting Kit 8(CCK-8) and Transwell assay were used to detect the proliferation and invasion of U87/TMZ cells. Dual luciferase reporter gene assay was used to verify the targeting relationship between miR-640 and SLIT1. Western blot was used to detect the expression levels of Wnt/β-catenin signaling pathway related proteins. [Results] miR-640 was highly expressed in glioma cells. The miR-640 expression level, number of invaded cells(57±23 vs 153±31), IC50 value(131.61±8.97 vs 293.33±11.28) and proliferation rate(18.48±4.23 vs 43.84±8.94) in miR-640 inhibitor group were significantly lower than those in NC inhibitor group(P<0.05). The miR-640 specifically regulated the expression of SLIT1, and the protein expression levels of β-catenin, c-myc and Cyclin D1 in miR-640 inhibitor+ si-SLIT1 group were higher than those in miR-640 inhibitor group(P<0.05). [Conclusion] miR-640 may promote glioma proliferation, invasion and resistance to TMZ through SLIT1-mediated Wnt/β-catenin pathway.
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