目的探讨外周血miR-141、miR-451a与弥漫大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)患者化疗应答的预测价值。方法选取2021年6月至2023年4月我院92例DLBCL患者作为DLBCL组,根据化疗效果分为化疗无效亚组(n=29)与化疗有效亚组(...目的探讨外周血miR-141、miR-451a与弥漫大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)患者化疗应答的预测价值。方法选取2021年6月至2023年4月我院92例DLBCL患者作为DLBCL组,根据化疗效果分为化疗无效亚组(n=29)与化疗有效亚组(n=63)。随机选取同期92例入院体检健康者为对照组,采用实时荧光定量聚合酶链反应测定miR-141、miR-451a相对表达量。比较DLBCL组与健康对照组外周血miR-141、miR-451a表达,以logistic回归模型分析筛选DLBCL患者化疗应答影响因素,相关性分析DLBCL患者外周血miR-141、miR-451a与国际预后指数(international prognositic index,IPI)评分、Ann Arbor分期间相关性,受试者工作特征(receiver operating characteristic,ROC)曲线评价DLBCL患者miR-141、miR-451a单项检测及联合检测预测化疗应答的价值。结果DLBCL患者外周血miR-141、miR-451a表达均低于健康对照组(P<0.05);logistic回归分析结果显示Ann Arbor分期、IPI评分均为DLBCL患者化疗应答独立危险因素,外周血miR-141、miR-451a均为DLBCL患者化疗应答性独立保护因素(P<0.05);DLBCL患者外周血miR-141、miR-451a与IPI评分、Ann Arbor分期均具有显著负相关关系(P<0.05);外周血miR-141、miR-451a单独预测DLBCL患者化疗应答的曲线下面积(area under the curve,AUC)值分别为0.770、0.794,二者联合预测AUC值高达0.929,明显高于miR-141、miR-451a单独预测,此时灵敏度、特异度分别为86.21%、85.71%。结论DCBCL患者血清miR-141、miR-451a表达下调,且与应答有关,检测二者水平,可预测DCBCL患者化疗应答,为临床工作提供参考。展开更多
目的:基于miR-451a靶向磷酸甘油酸变位酶(PGAM5)-线粒体动力相关蛋白1(Drp1)轴调控线粒体动力学,探讨温肺降浊方对氧糖剥夺/复氧(OGD/R)诱导HT22模型的效应机制。方法:建立OGD/R诱导HT22细胞模型,设立正常对照组、OGD/R组、miR-451a过...目的:基于miR-451a靶向磷酸甘油酸变位酶(PGAM5)-线粒体动力相关蛋白1(Drp1)轴调控线粒体动力学,探讨温肺降浊方对氧糖剥夺/复氧(OGD/R)诱导HT22模型的效应机制。方法:建立OGD/R诱导HT22细胞模型,设立正常对照组、OGD/R组、miR-451a过表达组、miR-451a敲低组、温肺降浊方含药血清(WFJZF)组、miR-451a过表达+WFJZF组、miR-451a敲低+WFJZF组和miR-451a空载体组,双荧光素酶确定miR-451a和PGAM5的转录关系,免疫蛋白共沉淀确定PGAM5和Drp1的调控作用,慢病毒转染miR-451a于HT22细胞中,qRT-PCR确定转染效率,CCK-8测定缺糖缺氧状态下细胞活性的最佳时间点,qRT-PCR和Westem Blot检测HT22模型细胞内DGAM5-Drp1轴蛋白及mRNA的表达水平。结果:OGD/R诱导HT22细胞持续2 h为最佳时间点,miR-451a可以靶向调控PGAM5基因,PGAM5和Drp1具有相互调控作用。与正常对照组比较,OGD/R组细胞内PGAM5、Drp1和Fis1蛋白及mRNA表达上升,p-Drp1 Ser 616磷酸化表达上升(P<0.05),OPA1蛋白及mRNA表达下降,p-Drp1 Ser 637去磷酸化表达下降(P<0.05);与OGD/R组比较,miR-451a过表达+WFJZF组细胞内PGAM5、Drp1和Fis1蛋白及mRNA表达下降,p-Drp1 Ser 616磷酸化表达下降(P<0.05),OPA1蛋白及mRNA表达上升,p-Drp1 Ser 637去磷酸化表达上升(P<0.05)。结论:miR-451a可以靶向调控PGAM5-Drp1轴,miR-451a过表达+WFJZF组可以改善线粒体失衡状态,减少神经元过度损伤,发挥脑保护效应。展开更多
BACKGROUND Aberrant microRNAs expression and associated pathways have been proved participate in regulation vast various physiologic and pathologic processed of different human cancers including liver cancer.While,the...BACKGROUND Aberrant microRNAs expression and associated pathways have been proved participate in regulation vast various physiologic and pathologic processed of different human cancers including liver cancer.While,the function of miR-451a in liver cancer still indistinct.AIM To study the effect of miR-451a in liver cancer development.METHODS GeneChip microarray analysis performed to detect miR-451a expression in liver cancer tissues and normal liver tissues.Reverse transcription-polymerase chain reaction was used to validate the expression of miR-451a in liver cancer cell and other tumor cell lines.Construction of liver cancer cell lines that stably overexpressed miR-451a by transfecting Lentivirus produced by Genechem company.Methylthiazolyldiphenyl-tetrazolium(MTT)bromide assay and colony formation assay to determine the effect of miR-451a in liver cancer cell proliferation.Flow cytometry used to investigate whether miR-451a involved in liver cancer cell apoptosis.Cell migration ability was measured via wound scratch assay.Target gene was explored by bioinformatic analysis,and downstream molecule of miR-451a in liver cancer identified by rescue experiments.RESULTS MiR-451a expression significantly downregulation in liver cancer tissues compared with that in normal liver tissue.MiR-451a also obviously low-expressed in liver cancer cell,colorectal carcinoma cell and esophageal carcinoma cell lines.Human hepatoblastoma G2(HepG2)and BEL-7404 cell lines that stably overexpressed miR-451a by transfecting lentivirus constructed successfully.MTT bromide assay and colony formation assay showed that the overexpressed miR-451a inhibit HepG2 cell proliferation viability,but not BEL-7404 cell.Flow cytometry determined that miR-451a regulating proliferation not through inducing apoptosis.Wound scratch assay revealed that miR-451a overexpression suppressed HepG2 cell migration.Furthermore,mex-3 RNA binding family member C was predicted as the target gene by bioinformatic analysis,and rescue experiments confirmed the hypothesis.CONCLUSION Therefore,miR-451a may be candidate miRNA for understanding molecular mechanisms of liver cancer development and novel target in liver cancer cell.展开更多
文摘目的探讨外周血miR-141、miR-451a与弥漫大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)患者化疗应答的预测价值。方法选取2021年6月至2023年4月我院92例DLBCL患者作为DLBCL组,根据化疗效果分为化疗无效亚组(n=29)与化疗有效亚组(n=63)。随机选取同期92例入院体检健康者为对照组,采用实时荧光定量聚合酶链反应测定miR-141、miR-451a相对表达量。比较DLBCL组与健康对照组外周血miR-141、miR-451a表达,以logistic回归模型分析筛选DLBCL患者化疗应答影响因素,相关性分析DLBCL患者外周血miR-141、miR-451a与国际预后指数(international prognositic index,IPI)评分、Ann Arbor分期间相关性,受试者工作特征(receiver operating characteristic,ROC)曲线评价DLBCL患者miR-141、miR-451a单项检测及联合检测预测化疗应答的价值。结果DLBCL患者外周血miR-141、miR-451a表达均低于健康对照组(P<0.05);logistic回归分析结果显示Ann Arbor分期、IPI评分均为DLBCL患者化疗应答独立危险因素,外周血miR-141、miR-451a均为DLBCL患者化疗应答性独立保护因素(P<0.05);DLBCL患者外周血miR-141、miR-451a与IPI评分、Ann Arbor分期均具有显著负相关关系(P<0.05);外周血miR-141、miR-451a单独预测DLBCL患者化疗应答的曲线下面积(area under the curve,AUC)值分别为0.770、0.794,二者联合预测AUC值高达0.929,明显高于miR-141、miR-451a单独预测,此时灵敏度、特异度分别为86.21%、85.71%。结论DCBCL患者血清miR-141、miR-451a表达下调,且与应答有关,检测二者水平,可预测DCBCL患者化疗应答,为临床工作提供参考。
文摘目的:基于miR-451a靶向磷酸甘油酸变位酶(PGAM5)-线粒体动力相关蛋白1(Drp1)轴调控线粒体动力学,探讨温肺降浊方对氧糖剥夺/复氧(OGD/R)诱导HT22模型的效应机制。方法:建立OGD/R诱导HT22细胞模型,设立正常对照组、OGD/R组、miR-451a过表达组、miR-451a敲低组、温肺降浊方含药血清(WFJZF)组、miR-451a过表达+WFJZF组、miR-451a敲低+WFJZF组和miR-451a空载体组,双荧光素酶确定miR-451a和PGAM5的转录关系,免疫蛋白共沉淀确定PGAM5和Drp1的调控作用,慢病毒转染miR-451a于HT22细胞中,qRT-PCR确定转染效率,CCK-8测定缺糖缺氧状态下细胞活性的最佳时间点,qRT-PCR和Westem Blot检测HT22模型细胞内DGAM5-Drp1轴蛋白及mRNA的表达水平。结果:OGD/R诱导HT22细胞持续2 h为最佳时间点,miR-451a可以靶向调控PGAM5基因,PGAM5和Drp1具有相互调控作用。与正常对照组比较,OGD/R组细胞内PGAM5、Drp1和Fis1蛋白及mRNA表达上升,p-Drp1 Ser 616磷酸化表达上升(P<0.05),OPA1蛋白及mRNA表达下降,p-Drp1 Ser 637去磷酸化表达下降(P<0.05);与OGD/R组比较,miR-451a过表达+WFJZF组细胞内PGAM5、Drp1和Fis1蛋白及mRNA表达下降,p-Drp1 Ser 616磷酸化表达下降(P<0.05),OPA1蛋白及mRNA表达上升,p-Drp1 Ser 637去磷酸化表达上升(P<0.05)。结论:miR-451a可以靶向调控PGAM5-Drp1轴,miR-451a过表达+WFJZF组可以改善线粒体失衡状态,减少神经元过度损伤,发挥脑保护效应。
基金Supported by National Natural Science Foundation of China,No.31640052and Natural Science Foundation of Shandong Province,China,No.ZR2019BH031.
文摘BACKGROUND Aberrant microRNAs expression and associated pathways have been proved participate in regulation vast various physiologic and pathologic processed of different human cancers including liver cancer.While,the function of miR-451a in liver cancer still indistinct.AIM To study the effect of miR-451a in liver cancer development.METHODS GeneChip microarray analysis performed to detect miR-451a expression in liver cancer tissues and normal liver tissues.Reverse transcription-polymerase chain reaction was used to validate the expression of miR-451a in liver cancer cell and other tumor cell lines.Construction of liver cancer cell lines that stably overexpressed miR-451a by transfecting Lentivirus produced by Genechem company.Methylthiazolyldiphenyl-tetrazolium(MTT)bromide assay and colony formation assay to determine the effect of miR-451a in liver cancer cell proliferation.Flow cytometry used to investigate whether miR-451a involved in liver cancer cell apoptosis.Cell migration ability was measured via wound scratch assay.Target gene was explored by bioinformatic analysis,and downstream molecule of miR-451a in liver cancer identified by rescue experiments.RESULTS MiR-451a expression significantly downregulation in liver cancer tissues compared with that in normal liver tissue.MiR-451a also obviously low-expressed in liver cancer cell,colorectal carcinoma cell and esophageal carcinoma cell lines.Human hepatoblastoma G2(HepG2)and BEL-7404 cell lines that stably overexpressed miR-451a by transfecting lentivirus constructed successfully.MTT bromide assay and colony formation assay showed that the overexpressed miR-451a inhibit HepG2 cell proliferation viability,but not BEL-7404 cell.Flow cytometry determined that miR-451a regulating proliferation not through inducing apoptosis.Wound scratch assay revealed that miR-451a overexpression suppressed HepG2 cell migration.Furthermore,mex-3 RNA binding family member C was predicted as the target gene by bioinformatic analysis,and rescue experiments confirmed the hypothesis.CONCLUSION Therefore,miR-451a may be candidate miRNA for understanding molecular mechanisms of liver cancer development and novel target in liver cancer cell.
