目的 探讨长链非编码RNA(long non-coding RNA,lncRNA)KCNIP4-IT1/miR-181c-5p分子轴对喉癌TU177细胞增殖和迁移的调控作用。方法 采用qPCR检测30对喉癌组织和癌旁组织、喉癌细胞系(TU159、TU686、AMC-NH-8、TU177、TU138)和支气管上皮...目的 探讨长链非编码RNA(long non-coding RNA,lncRNA)KCNIP4-IT1/miR-181c-5p分子轴对喉癌TU177细胞增殖和迁移的调控作用。方法 采用qPCR检测30对喉癌组织和癌旁组织、喉癌细胞系(TU159、TU686、AMC-NH-8、TU177、TU138)和支气管上皮细胞系(16HBE)中KCNIP4-IT1表达水平。通过RNA干扰技术将KCNIP4-IT1的小干扰RNA转染到TU177细胞(si-KCNIP4-IT1组),建立KCNIP4-IT1低表达细胞系,另设置阴性Ctrl组(Ctrl组,转染阴性对照序列)。MTT法、划痕愈合实验检测敲低KCNIP4-IT1后TU177细胞增殖和迁移的变化。双荧光素酶报告基因实验验证KCNIP4-IT1与miR-181c-5p之间的靶向调控关系。qPCR检测细胞中miR-181c-5p和人纤溶酶原激活物抑制剂1(SERPINE1)mRNA的表达水平。Western blotting检测细胞迁移蛋白(β-catenin、N-Cadherin)和增殖蛋白(CDK3、PCNA)的表达水平。结果 KCNIP4-IT1在喉癌组织中表达水平高于癌旁组织(t=9.297,P<0.01)。KCNIP4-IT1在喉癌细胞系中表达水平高于16HBE细胞(t分别为4.620、7.502、4.944、8.665、9.139,P均<0.01),以TU177细胞中KCNIP4-IT1表达水平最高(F=17.819,P <0.01)。与Ctrl组相比,敲低KCNIP4-IT1显著抑制TU177细胞增殖(P均<0.05)和迁移(t=3.740,P<0.01)。双荧光素酶报告基因实验证实KCNIP4-IT1靶向调控miR-181c-5p(t=5.838,P<0.01)。与Ctrl组相比,敲低KCNIP4-IT1明显上调miR-181c-5p的表达水平(t=6.214,P <0.01),下调SE R P I N E1基因的表达水平(t=8.19 0,P<0.01)。与Ctrl组相比,敲低KCNIP4-IT1明显下调细胞迁移蛋白(β-catenin、N-Cadherin)和增殖蛋白(CDK3、PCNA)的表达水平。结论 KCNIP4-IT1在喉癌组织和细胞系中呈高表达状态,敲低KCNIP4-IT1抑制喉癌TU177细胞的增殖和迁移,其分子机制可能与靶向调控miR-181c-5p并抑制SERPINE1表达有关。展开更多
Severe cerebral ischemia/reperfusion injury has been shown to induce high-level autophagy and neuronal death.Therefore,it is extremely important to search for a target that inhibits autophagy activation.Long non-codin...Severe cerebral ischemia/reperfusion injury has been shown to induce high-level autophagy and neuronal death.Therefore,it is extremely important to search for a target that inhibits autophagy activation.Long non-coding RNA MEG3 participates in autophagy.However,it remains unclear whether it can be targeted to regulate cerebral ischemia/reperfusion injury.Our results revealed that in oxygen and glucose deprivation/reoxygenation-treated HT22 cells,MEG3 expression was obviously upregulated,and autophagy was increased,while knockdown of MEG3 expression greatly reduced autophagy.Furthermore,MEG3 bound mi R-181 c-5 p and inhibited its expression,while mi R-181 c-5 p bound to autophagy-related gene ATG7 and inhibited its expression.Further experiments revealed that mir-181 c-5 p overexpression reversed the effect of MEG3 on autophagy and ATG7 expression in HT22 cells subjected to oxygen and glucose deprivation/reoxygenation.In vivo experiments revealed that MEG3 knockdown suppressed autophagy,infarct volume and behavioral deficits in cerebral ischemia/reperfusion mice.These findings suggest that MEG3 knockdown inhibited autophagy and alleviated cerebral ischemia/reperfusion injury through the mi R-181 c-5 p/ATG7 signaling pathway.Therefore,MEG3 can be considered as an intervention target for the treatment of cerebral ischemia/reperfusion injury.This study was approved by the Animal Ethics Committee of the First Affiliated Hospital of Zhengzhou University,China(approval No.XF20190538)on January 4,2019.展开更多
Objective To preliminarily explore the effects of human microRNA-181a on migration of gastric cancer cells and its mechanism.Methods The expression of miRNA-181a-5p in gastric cancer cell line GC9811 and peritoneal hi...Objective To preliminarily explore the effects of human microRNA-181a on migration of gastric cancer cells and its mechanism.Methods The expression of miRNA-181a-5p in gastric cancer cell line GC9811 and peritoneal high metastasis gastric cancer cell line GC9811-P were tested by quantitative real-time polymerase chain reaction(qRT-PCR).GC9811 cell line was展开更多
文摘目的 探讨长链非编码RNA(long non-coding RNA,lncRNA)KCNIP4-IT1/miR-181c-5p分子轴对喉癌TU177细胞增殖和迁移的调控作用。方法 采用qPCR检测30对喉癌组织和癌旁组织、喉癌细胞系(TU159、TU686、AMC-NH-8、TU177、TU138)和支气管上皮细胞系(16HBE)中KCNIP4-IT1表达水平。通过RNA干扰技术将KCNIP4-IT1的小干扰RNA转染到TU177细胞(si-KCNIP4-IT1组),建立KCNIP4-IT1低表达细胞系,另设置阴性Ctrl组(Ctrl组,转染阴性对照序列)。MTT法、划痕愈合实验检测敲低KCNIP4-IT1后TU177细胞增殖和迁移的变化。双荧光素酶报告基因实验验证KCNIP4-IT1与miR-181c-5p之间的靶向调控关系。qPCR检测细胞中miR-181c-5p和人纤溶酶原激活物抑制剂1(SERPINE1)mRNA的表达水平。Western blotting检测细胞迁移蛋白(β-catenin、N-Cadherin)和增殖蛋白(CDK3、PCNA)的表达水平。结果 KCNIP4-IT1在喉癌组织中表达水平高于癌旁组织(t=9.297,P<0.01)。KCNIP4-IT1在喉癌细胞系中表达水平高于16HBE细胞(t分别为4.620、7.502、4.944、8.665、9.139,P均<0.01),以TU177细胞中KCNIP4-IT1表达水平最高(F=17.819,P <0.01)。与Ctrl组相比,敲低KCNIP4-IT1显著抑制TU177细胞增殖(P均<0.05)和迁移(t=3.740,P<0.01)。双荧光素酶报告基因实验证实KCNIP4-IT1靶向调控miR-181c-5p(t=5.838,P<0.01)。与Ctrl组相比,敲低KCNIP4-IT1明显上调miR-181c-5p的表达水平(t=6.214,P <0.01),下调SE R P I N E1基因的表达水平(t=8.19 0,P<0.01)。与Ctrl组相比,敲低KCNIP4-IT1明显下调细胞迁移蛋白(β-catenin、N-Cadherin)和增殖蛋白(CDK3、PCNA)的表达水平。结论 KCNIP4-IT1在喉癌组织和细胞系中呈高表达状态,敲低KCNIP4-IT1抑制喉癌TU177细胞的增殖和迁移,其分子机制可能与靶向调控miR-181c-5p并抑制SERPINE1表达有关。
文摘Severe cerebral ischemia/reperfusion injury has been shown to induce high-level autophagy and neuronal death.Therefore,it is extremely important to search for a target that inhibits autophagy activation.Long non-coding RNA MEG3 participates in autophagy.However,it remains unclear whether it can be targeted to regulate cerebral ischemia/reperfusion injury.Our results revealed that in oxygen and glucose deprivation/reoxygenation-treated HT22 cells,MEG3 expression was obviously upregulated,and autophagy was increased,while knockdown of MEG3 expression greatly reduced autophagy.Furthermore,MEG3 bound mi R-181 c-5 p and inhibited its expression,while mi R-181 c-5 p bound to autophagy-related gene ATG7 and inhibited its expression.Further experiments revealed that mir-181 c-5 p overexpression reversed the effect of MEG3 on autophagy and ATG7 expression in HT22 cells subjected to oxygen and glucose deprivation/reoxygenation.In vivo experiments revealed that MEG3 knockdown suppressed autophagy,infarct volume and behavioral deficits in cerebral ischemia/reperfusion mice.These findings suggest that MEG3 knockdown inhibited autophagy and alleviated cerebral ischemia/reperfusion injury through the mi R-181 c-5 p/ATG7 signaling pathway.Therefore,MEG3 can be considered as an intervention target for the treatment of cerebral ischemia/reperfusion injury.This study was approved by the Animal Ethics Committee of the First Affiliated Hospital of Zhengzhou University,China(approval No.XF20190538)on January 4,2019.
文摘Objective To preliminarily explore the effects of human microRNA-181a on migration of gastric cancer cells and its mechanism.Methods The expression of miRNA-181a-5p in gastric cancer cell line GC9811 and peritoneal high metastasis gastric cancer cell line GC9811-P were tested by quantitative real-time polymerase chain reaction(qRT-PCR).GC9811 cell line was
