Background:Glioblastoma remains a highly invasive primary brain malignancy with an undesirable prognosis.Growing evidence has shed light on the importance of microRNAs(miRs),as small non-coding RNAs,in tumor developme...Background:Glioblastoma remains a highly invasive primary brain malignancy with an undesirable prognosis.Growing evidence has shed light on the importance of microRNAs(miRs),as small non-coding RNAs,in tumor development and progression.The present study leverages the in-silico and in-vitro techniques to investigate the significance of hsa-miR-181a-5p and the underlying hsa-miR-181a-5p-meidated signaling pathway in glioblastoma development.Methods:Bioinformatic studies were performed on GSE158284,GSE108474(REMBRANDT study),TCGA-GTEx,CCLE,GeneMANIA,Reactome,WikiPathways,KEGG,miRDB,and microT-CDS to identify the significance of hsa-miR-181a-5p and its underlying target.Afterward,the U373 cell line was selected and transfected with hsa-miR-181a-5p mimics,and the cell viability,clonogenicity,migration,mRNA expression,apoptosis,and cell cycle were studied using the MTT assay,colony formation test,migration assay,qRT-PCR,andflow cytometry respectively.Results:hsa-miR-181a-5p expression is decreased in glioblastoma samples.The in-silico results have shown that hsa-miR-181a-5p could regulate the MAPK pathway by targeting AKT3.The experimental assays have shown that hsa-miR-181a-5p decreases the migration of glioblastoma cells,arrests the cell cycle,and increases the apoptosis rate.Besides downregulating MMP9 and upregulating BAX,hsa-miR-181a-5p downregulates MET,MAP2K1,MAPK1,MAPK3,and AKT3 expression in U373 cells.The in-vitro results were consistent with in-silico results regarding the regulatory effect of hsa-miR-181a-5p on the MAPK pathway,leading to tumor suppression in glioblastoma.Conclusions:hsa-miR-181a-5p inhibits glioblastoma development partially by regulating the signaling factors of the MAPK pathway.展开更多
目的观察hsa-miR-93对肝癌细胞生长和细胞周期的影响,以分析其在肝癌中的作用机制。方法用real time RT-PCR法分析hsa-miR-93在人肝癌和癌旁肝组织的表达情况。此后构建hsa-miR-93过表达载体,同时合成二甲氧修饰的hsa-miR-93的反义核苷...目的观察hsa-miR-93对肝癌细胞生长和细胞周期的影响,以分析其在肝癌中的作用机制。方法用real time RT-PCR法分析hsa-miR-93在人肝癌和癌旁肝组织的表达情况。此后构建hsa-miR-93过表达载体,同时合成二甲氧修饰的hsa-miR-93的反义核苷酸序列,二者转染肝癌细胞株观察它们对肝癌细胞生长及细胞周期的影响,分别以空载体和无关寡核苷酸序列为对照。转染后CCK-8法检测肝癌细胞生长情况,通过流式细胞仪分析细胞周期。结果hsa-miR-93在肝癌组织中表达明显高于癌旁肝组织。hsa-miR-93过表达载体可以促进肝癌细胞生长,促进细胞周期的G1/S期转换,而hsa-miR-93反义序列抑制肝癌细胞生长,使肝癌细胞阻滞于G1期(P<0.05)。结论hsa-miR-93通过促进细胞周期的G1/S期转换而促进肝癌细胞生长,提示hsa-miR-93可能是肝癌的发病机制中一个重要分子。展开更多
文摘Background:Glioblastoma remains a highly invasive primary brain malignancy with an undesirable prognosis.Growing evidence has shed light on the importance of microRNAs(miRs),as small non-coding RNAs,in tumor development and progression.The present study leverages the in-silico and in-vitro techniques to investigate the significance of hsa-miR-181a-5p and the underlying hsa-miR-181a-5p-meidated signaling pathway in glioblastoma development.Methods:Bioinformatic studies were performed on GSE158284,GSE108474(REMBRANDT study),TCGA-GTEx,CCLE,GeneMANIA,Reactome,WikiPathways,KEGG,miRDB,and microT-CDS to identify the significance of hsa-miR-181a-5p and its underlying target.Afterward,the U373 cell line was selected and transfected with hsa-miR-181a-5p mimics,and the cell viability,clonogenicity,migration,mRNA expression,apoptosis,and cell cycle were studied using the MTT assay,colony formation test,migration assay,qRT-PCR,andflow cytometry respectively.Results:hsa-miR-181a-5p expression is decreased in glioblastoma samples.The in-silico results have shown that hsa-miR-181a-5p could regulate the MAPK pathway by targeting AKT3.The experimental assays have shown that hsa-miR-181a-5p decreases the migration of glioblastoma cells,arrests the cell cycle,and increases the apoptosis rate.Besides downregulating MMP9 and upregulating BAX,hsa-miR-181a-5p downregulates MET,MAP2K1,MAPK1,MAPK3,and AKT3 expression in U373 cells.The in-vitro results were consistent with in-silico results regarding the regulatory effect of hsa-miR-181a-5p on the MAPK pathway,leading to tumor suppression in glioblastoma.Conclusions:hsa-miR-181a-5p inhibits glioblastoma development partially by regulating the signaling factors of the MAPK pathway.
文摘目的观察hsa-miR-93对肝癌细胞生长和细胞周期的影响,以分析其在肝癌中的作用机制。方法用real time RT-PCR法分析hsa-miR-93在人肝癌和癌旁肝组织的表达情况。此后构建hsa-miR-93过表达载体,同时合成二甲氧修饰的hsa-miR-93的反义核苷酸序列,二者转染肝癌细胞株观察它们对肝癌细胞生长及细胞周期的影响,分别以空载体和无关寡核苷酸序列为对照。转染后CCK-8法检测肝癌细胞生长情况,通过流式细胞仪分析细胞周期。结果hsa-miR-93在肝癌组织中表达明显高于癌旁肝组织。hsa-miR-93过表达载体可以促进肝癌细胞生长,促进细胞周期的G1/S期转换,而hsa-miR-93反义序列抑制肝癌细胞生长,使肝癌细胞阻滞于G1期(P<0.05)。结论hsa-miR-93通过促进细胞周期的G1/S期转换而促进肝癌细胞生长,提示hsa-miR-93可能是肝癌的发病机制中一个重要分子。
