摘要
目的探讨微小RNA-524-5p(miR-524-5p)调控叉头框D3蛋白(FOXD3)对甲状腺癌细胞增殖、凋亡及上皮间质转化(EMT)的影响。方法人甲状腺癌细胞SW579,分为空白对照组(不转染)、mimic NC组(转染negative control-miR-524-5p)、miR-524-5p mimic组(转染hsa-miR-524-5p mimic)、siRNA NC组(同时转染hsa-miR-524-5p mimic和NC-siRNA)、共转染组(同时转染hsa-miR-524-5p mimic和FOXD3 siRNA),实时荧光定量PCR(qRT-PCR)法检测miR-524-5p、FOXD3 mRNA表达;CCK-8法、流式细胞仪检测细胞增殖、凋亡;Transwell法、划痕试验检测细胞侵袭、迁移能力;蛋白印迹(WB)法检测各组SW579细胞中FOXD3及EMT相关因子E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、纤连蛋白(Fibronectin)蛋白表达情况。结果与mimic NC组相比,miR-524-5p mimic组SW579细胞中miR-524-5p表达水平、细胞凋亡率、FOXD3 mRNA和蛋白表达水平、E-cadherin蛋白表达水平显著升高(P<0.05),细胞OD值、侵袭细胞数、细胞迁移率、N-cadherin、Vimentin、Fibronectin蛋白表达水平显著降低(P<0.05);与siRNA NC组相比,共转染组SW579细胞中miR-524-5p表达水平差异无统计学意义(P>0.05),细胞OD值、侵袭细胞数、细胞迁移率、N-cadherin、Vimentin、Fibronectin蛋白表达水平显著升高(P<0.05),细胞凋亡率、FOXD3 mRNA和蛋白表达水平、E-cadherin蛋白表达水平显著降低(P>0.05)。结论miR-524-5p可能通过调控FOXD3表达水平升高,进而抑制SW579细胞增殖、侵袭、迁移及EMT过程,并诱导癌细胞凋亡。
Objective To investigate the effects of microRNA-524-5p(miR-524-5p)regulating forkhead box D3(FOXD3)on proliferation,apoptosis and epithelial mesenchymal transition(EMT)of thyroid cancer cells.Methods SW579 cells were cultured in vitro and divided into blank control group(without transfection),mimic NC group(transfected with negative control-miR-524-5p),miR-524-5p mimic group(transfected with hsa-miR-524-5p mimic),siRNA NC group(transfected with hsa-miR-524-5p mimic and NC-siRNA)and co-transfection group(transfected with hsa-miR-524-5p mimic and FOXD3 siRNA).The expressions of miR-524-5p and FOXD3 mRNA were assessed by real-time fluorescence quantitative PCR(qRT-PCR).The proliferation and apoptosis of cells was detected by CCK-8 method and flow cytometry.The invasion and migration ability of cells was detected by transwell method and scratch test.In addition,the protein expressions of FOXD3 and EMT related factors E-cadherin,N-cadherin,Vimentin and Fibronectin in SW579 cells were detected by Western blot(WB).Results Compared with those in mimic NC group,miR-524-5p expression level,apoptosis rate,FOXD3 mRNA,FOXD3 protein and E-cadherin protein levels were significantly higher in miR-524-5p mimic group(P<0.05);and OD value,invasive cells number,migration rate,N-cadherin,vimentin and fibronectin protein expression levels were significantly lower(P<0.05).Compared with that in siRNA NC group,there was no significant difference in miR-524-5p expression level in SW579 cells in co-transfection group(P>0.05);while OD value,invasive cells number,migration rate,N-cadherin,vimentin and fibronectin protein expression levels were significantly higher(P<0.05),and the apoptosis rate,FOXD3 mRNA,FOXD3 protein and E-cadherin protein levels were significantly lower(P<0.05).Conclusion MiR-524-5p may inhibit the proliferation,invasion,migration and EMT process of SW579 cells by up-regulating the expression of FOXD3,so as to induce the apoptosis of SW579 cells.
作者
殷鹏昌
赵良柱
闫红印
YIN Peng-chang;ZHAO Liang-zhu;YAN Hong-yin(Department of General Surgery,Pingmei Shenma Group General Hospital,Pingdingshan 461700,Henan,China;不详)
出处
《广东医学》
CAS
2022年第7期860-865,共6页
Guangdong Medical Journal
基金
河南省医学科技公关计划联合共建项目(LHGJ20190028)。
