BACKGROUND Ectopic expression of miRNAs promotes tumor development and progression.miRNA(miR)-320a is downregulated in many cancers,including gastric cancer(GC).However,the mechanism underlying its downregulation and ...BACKGROUND Ectopic expression of miRNAs promotes tumor development and progression.miRNA(miR)-320a is downregulated in many cancers,including gastric cancer(GC).However,the mechanism underlying its downregulation and the role of miR-320a in GC are unknown.AIM To determine expression and biological functions of miR-320a in GC and investigate the underlying molecular mechanisms.METHODS Quantitative real-time polymerase chain reaction(PCR)was used to determine expression of miR-320a in GC cell lines and tissues.TargetScanHuman7.1,miRDB,and microRNA.org were used to predict the possible targets of miR-320a,and a dual luciferase assay was used to confirm the findings.Western blotting was used to detect the protein levels of pre-B-cell leukemia homeobox 3(PBX3)in GC cells and tissue samples.Cell Counting Kit-8 proliferation,Transwell,wound healing,and apoptosis assays were performed to analyze the biological functions of miR-320a in GC cells.Methylation-specific PCR was used to analyze the methylation level of the miR-320a promoter CpG islands.5-Aza-2’-deoxycytidine(5-Aza-CdR)and trichostatin A(TSA)were used to treat GC cells.RESULTS miR-320a expression was lower in GC cell lines and tissues than in the normal gastric mucosa cell line GES-1 and matched adjacent normal tissues.miR-320a overexpression suppressed GC cell proliferation,invasion and migration,and induced apoptosis.PBX3 was a target of miR-320a in GC.The methylation level of the miR-320a promoter CpG islands was elevated and this was partly reversed by 5-Aza-CdR and TSA.CONCLUSION miR-320a acts as a tumor suppressor and inhibits malignant behavior of GC cells,partly by targeting PBX3.DNA methylation is an important mechanism associated with low expression of miR-320a.展开更多
We rechecked the original data of Figure 3,Part.B,and found that 0 h group in the BGC-823 cell wound scratch assay was misapplied.Therefore,we are writing to apply for the modification of Figure 3,Part.B.
BACKGROUND Necrotizing enterocolitis(NEC)remains a prominent gastrointestinal emergency among infants,particularly term infants with congenital heart defects(CHD)being at high risk.The molecular processes that contrib...BACKGROUND Necrotizing enterocolitis(NEC)remains a prominent gastrointestinal emergency among infants,particularly term infants with congenital heart defects(CHD)being at high risk.The molecular processes that contribute to NEC have yet to be completely understood.The high mortality rates necessitate an active search for noninvasive biomarkers that can aid in the preclinical diagnosis and prognosis of NEC.MicroRNAs(miRs),which are involved in many biological processes in both health and disease,have been discovered to play an important role in regulating inflammation and immune responses via various signaling pathways.AIM To determine the plasma levels of miR-155,miR-221,miR-223,miR-320a,miR-451a as potential NEC biomarkers in term newborns with CHD.METHODS This prospective cohort study included twenty-tree term newborns with CHD who underwent cardiac surgery on the median day of life(DOL)=7.Nine of them developed NEC(Bell’s stage IIA and IIIA)within 1 week of cardiac surgery(NEC newborns).Blood samples were collected before(median DOL=5)and following(median DOL=13)cardiac surgery.Levels of plasma miR-155-5p,miR-221-3p,miR-223-3p,miR-320a-3p,and miR-451a were determined using real-time polymerase chain reaction.The functional analysis was executed using the DIANA-miRPath v4.0.RESULTS Preoperatively,NEC newborns had significantly lower plasma levels of miR-155(2.70-fold,P=0.020),miR-223(2.42-fold,P=0.030),and miR-320a(3.62-fold,P=0.006)than newborns without NEC.Postoperatively,miR-451a levels differed significantly between the newborn groups,showing a 4.70-fold decrease(P=0.014)in expression when clinical NEC symptoms appeared.According to receiver operating characteristic analysis,miR-320a was found to be the most effective predictive biomarker for NEC[area under the curve(AUC)=0.835,63%sensitivity,100%specificity],while miR-451a was identified as a NEC biomarker(AUC=0.835,85.7%sensitivity,76.9%specificity).Preoperatively,miR-155-5p,miR-223-3p,and miR-320a-3p were differentially expressed and targeted the forkhead box O and Hippo pathways(P<0.01).CONCLUSION Our study demonstrates,for the first time,that plasma miR-320a-3p levels can be used as a preclinical biomarker for NEC in term newborns with CHD.展开更多
目的:构建中草药来源的目标微小RNA(microRNA,miR)在小鼠肺组织中的靶基因谱检测方法,利用该方法检测清肺排毒汤所含miR-320—靶基因谱及其生物学功能,为清肺排毒汤治疗病毒性肺炎的分子机制补充miR维度的数据支撑。方法:采用二代高通...目的:构建中草药来源的目标微小RNA(microRNA,miR)在小鼠肺组织中的靶基因谱检测方法,利用该方法检测清肺排毒汤所含miR-320—靶基因谱及其生物学功能,为清肺排毒汤治疗病毒性肺炎的分子机制补充miR维度的数据支撑。方法:采用二代高通量测序测定目标miR(miR-320)在清肺排毒汤中的表达情况,利用“结合态miR—靶基因抓取测序技术(capturing and sequencing of miRNA-target complex technology,CSCT)”初步检测其在小鼠肺组织中的靶基因谱,并采用Alphafold3验证检测结果,取其交集作为目标miR的靶基因谱;进而分析靶基因谱的功能,阐释清肺排毒汤通过miR-320在小鼠肺组织中的调控功能;此外,将检测结果与TargetScan的预测结果进行比对,验证本检测方法(CSCT+Alphafold3序贯检测法)的优势。结果:高通量测序结果表明清肺排毒汤中富含miR-320,其表达含量居前50位。miR-320在小鼠肺组织中可作用于26类靶基因,其中19类为已知基因(18类为mRNA以及1类为转录增强子),其主要通过miR经典作用模式识别靶基因,与26类靶基因具有良好的碱基互补性,最小自由能在-35.8~-21.8 kcal·mol^(-1)之间;Alphafold3预测的靶基因的iPTM和PTM之和最小值为1.1,位于高置信度区域,100%验证了CSCT的检测结果,据此确证26类靶基因为miR-320作用谱;与TargetScan相比,本研究构建的“CSCT+Alphafold3序贯检测法”对于靶基因谱的检测准确率更高。这些靶基因具有多种功能,主要富集于白介素介导的免疫信号通路,在抗原处理与提呈、免疫因子或细胞介导的细胞凋亡、淋巴细胞增殖与活化等生物学过程中发挥作用。结论:清肺排毒汤所含miR-320在小鼠肺组织内主要富集于免疫调节相关通路,这可能是清肺排毒汤治疗病毒性肺炎的miR分子机制之一;本研究构建的“CSCT+Alphafold3”法对于miR靶基因谱的检测具有较高的可靠性和准确性,可作为中药汤剂中miR—靶基因互作谱检测技术。展开更多
【目的】研究microRNA-320d(miR-320d)对子宫内膜癌JEC细胞上皮-间质转化功能的影响及其相关机制。【方法】JEC子宫内膜癌细胞株分别转染miR-320d mimics和negative control mimic,分别作为M320d、NCM组,并设立未转染对照control组,采用...【目的】研究microRNA-320d(miR-320d)对子宫内膜癌JEC细胞上皮-间质转化功能的影响及其相关机制。【方法】JEC子宫内膜癌细胞株分别转染miR-320d mimics和negative control mimic,分别作为M320d、NCM组,并设立未转染对照control组,采用RT-PCR法检测各组细胞miR-320d含量。Transwell实验检测3组细胞迁移能力和侵袭能力。Westernblot法检测3组细胞α-Catenin和PBX3表达水平,及PBX3过表达对miR-320d抑制EMT的拮抗作用。双荧光素酶实验检测miR-320d与PBX3的关系。【结果】M320d组miR-320d的表达水平为control组的(808.25±15.58)倍(P<0.05)。M320d组迁移细胞数量29.56±0.59低于control组的94.48±1.02(P<0.05)。M320d组侵袭细胞数量7.33±0.84低于control组的86.28±3.51(P<0.05)。M320d组细胞α-Catenin、E-cadherin蛋白表达量(0.365±0.017、0.261±0.008)高于对照组(0.114±0.010、0.151±0.021),Vimentin蛋白表达量(0.105±0.009)低于对照组(0.211±0.025),PBX3蛋白表达量(0.181±0.002)低于对照组(0.311±0.007)。PBX3过表达M320d组子宫内膜癌细胞中α-Catenin、E-cadherin蛋白表达量(0.139±0.019、0.143±0.007)低于对照组(0.399±0.034、0.261±0.017),Vimentin蛋白表达量(0.234±0.008)高于对照组(0.105±0.009),差异均有统计学意义(P<0.05)。双荧光素酶检验结果显示PBX3为miR-320d的下游靶基因。【结论】miR-320d可能通过降低下游靶基因PBX3水平影响EMT相关蛋白表达,抑制子宫内膜癌JEC细胞的上皮-间质转化功能。展开更多
基金Supported by the Natural Science Foundation of Liaoning Province,No.201602817
文摘BACKGROUND Ectopic expression of miRNAs promotes tumor development and progression.miRNA(miR)-320a is downregulated in many cancers,including gastric cancer(GC).However,the mechanism underlying its downregulation and the role of miR-320a in GC are unknown.AIM To determine expression and biological functions of miR-320a in GC and investigate the underlying molecular mechanisms.METHODS Quantitative real-time polymerase chain reaction(PCR)was used to determine expression of miR-320a in GC cell lines and tissues.TargetScanHuman7.1,miRDB,and microRNA.org were used to predict the possible targets of miR-320a,and a dual luciferase assay was used to confirm the findings.Western blotting was used to detect the protein levels of pre-B-cell leukemia homeobox 3(PBX3)in GC cells and tissue samples.Cell Counting Kit-8 proliferation,Transwell,wound healing,and apoptosis assays were performed to analyze the biological functions of miR-320a in GC cells.Methylation-specific PCR was used to analyze the methylation level of the miR-320a promoter CpG islands.5-Aza-2’-deoxycytidine(5-Aza-CdR)and trichostatin A(TSA)were used to treat GC cells.RESULTS miR-320a expression was lower in GC cell lines and tissues than in the normal gastric mucosa cell line GES-1 and matched adjacent normal tissues.miR-320a overexpression suppressed GC cell proliferation,invasion and migration,and induced apoptosis.PBX3 was a target of miR-320a in GC.The methylation level of the miR-320a promoter CpG islands was elevated and this was partly reversed by 5-Aza-CdR and TSA.CONCLUSION miR-320a acts as a tumor suppressor and inhibits malignant behavior of GC cells,partly by targeting PBX3.DNA methylation is an important mechanism associated with low expression of miR-320a.
文摘We rechecked the original data of Figure 3,Part.B,and found that 0 h group in the BGC-823 cell wound scratch assay was misapplied.Therefore,we are writing to apply for the modification of Figure 3,Part.B.
基金Supported by The Russian Science Foundation,No.19-75-20076.
文摘BACKGROUND Necrotizing enterocolitis(NEC)remains a prominent gastrointestinal emergency among infants,particularly term infants with congenital heart defects(CHD)being at high risk.The molecular processes that contribute to NEC have yet to be completely understood.The high mortality rates necessitate an active search for noninvasive biomarkers that can aid in the preclinical diagnosis and prognosis of NEC.MicroRNAs(miRs),which are involved in many biological processes in both health and disease,have been discovered to play an important role in regulating inflammation and immune responses via various signaling pathways.AIM To determine the plasma levels of miR-155,miR-221,miR-223,miR-320a,miR-451a as potential NEC biomarkers in term newborns with CHD.METHODS This prospective cohort study included twenty-tree term newborns with CHD who underwent cardiac surgery on the median day of life(DOL)=7.Nine of them developed NEC(Bell’s stage IIA and IIIA)within 1 week of cardiac surgery(NEC newborns).Blood samples were collected before(median DOL=5)and following(median DOL=13)cardiac surgery.Levels of plasma miR-155-5p,miR-221-3p,miR-223-3p,miR-320a-3p,and miR-451a were determined using real-time polymerase chain reaction.The functional analysis was executed using the DIANA-miRPath v4.0.RESULTS Preoperatively,NEC newborns had significantly lower plasma levels of miR-155(2.70-fold,P=0.020),miR-223(2.42-fold,P=0.030),and miR-320a(3.62-fold,P=0.006)than newborns without NEC.Postoperatively,miR-451a levels differed significantly between the newborn groups,showing a 4.70-fold decrease(P=0.014)in expression when clinical NEC symptoms appeared.According to receiver operating characteristic analysis,miR-320a was found to be the most effective predictive biomarker for NEC[area under the curve(AUC)=0.835,63%sensitivity,100%specificity],while miR-451a was identified as a NEC biomarker(AUC=0.835,85.7%sensitivity,76.9%specificity).Preoperatively,miR-155-5p,miR-223-3p,and miR-320a-3p were differentially expressed and targeted the forkhead box O and Hippo pathways(P<0.01).CONCLUSION Our study demonstrates,for the first time,that plasma miR-320a-3p levels can be used as a preclinical biomarker for NEC in term newborns with CHD.
文摘目的:构建中草药来源的目标微小RNA(microRNA,miR)在小鼠肺组织中的靶基因谱检测方法,利用该方法检测清肺排毒汤所含miR-320—靶基因谱及其生物学功能,为清肺排毒汤治疗病毒性肺炎的分子机制补充miR维度的数据支撑。方法:采用二代高通量测序测定目标miR(miR-320)在清肺排毒汤中的表达情况,利用“结合态miR—靶基因抓取测序技术(capturing and sequencing of miRNA-target complex technology,CSCT)”初步检测其在小鼠肺组织中的靶基因谱,并采用Alphafold3验证检测结果,取其交集作为目标miR的靶基因谱;进而分析靶基因谱的功能,阐释清肺排毒汤通过miR-320在小鼠肺组织中的调控功能;此外,将检测结果与TargetScan的预测结果进行比对,验证本检测方法(CSCT+Alphafold3序贯检测法)的优势。结果:高通量测序结果表明清肺排毒汤中富含miR-320,其表达含量居前50位。miR-320在小鼠肺组织中可作用于26类靶基因,其中19类为已知基因(18类为mRNA以及1类为转录增强子),其主要通过miR经典作用模式识别靶基因,与26类靶基因具有良好的碱基互补性,最小自由能在-35.8~-21.8 kcal·mol^(-1)之间;Alphafold3预测的靶基因的iPTM和PTM之和最小值为1.1,位于高置信度区域,100%验证了CSCT的检测结果,据此确证26类靶基因为miR-320作用谱;与TargetScan相比,本研究构建的“CSCT+Alphafold3序贯检测法”对于靶基因谱的检测准确率更高。这些靶基因具有多种功能,主要富集于白介素介导的免疫信号通路,在抗原处理与提呈、免疫因子或细胞介导的细胞凋亡、淋巴细胞增殖与活化等生物学过程中发挥作用。结论:清肺排毒汤所含miR-320在小鼠肺组织内主要富集于免疫调节相关通路,这可能是清肺排毒汤治疗病毒性肺炎的miR分子机制之一;本研究构建的“CSCT+Alphafold3”法对于miR靶基因谱的检测具有较高的可靠性和准确性,可作为中药汤剂中miR—靶基因互作谱检测技术。
文摘【目的】研究microRNA-320d(miR-320d)对子宫内膜癌JEC细胞上皮-间质转化功能的影响及其相关机制。【方法】JEC子宫内膜癌细胞株分别转染miR-320d mimics和negative control mimic,分别作为M320d、NCM组,并设立未转染对照control组,采用RT-PCR法检测各组细胞miR-320d含量。Transwell实验检测3组细胞迁移能力和侵袭能力。Westernblot法检测3组细胞α-Catenin和PBX3表达水平,及PBX3过表达对miR-320d抑制EMT的拮抗作用。双荧光素酶实验检测miR-320d与PBX3的关系。【结果】M320d组miR-320d的表达水平为control组的(808.25±15.58)倍(P<0.05)。M320d组迁移细胞数量29.56±0.59低于control组的94.48±1.02(P<0.05)。M320d组侵袭细胞数量7.33±0.84低于control组的86.28±3.51(P<0.05)。M320d组细胞α-Catenin、E-cadherin蛋白表达量(0.365±0.017、0.261±0.008)高于对照组(0.114±0.010、0.151±0.021),Vimentin蛋白表达量(0.105±0.009)低于对照组(0.211±0.025),PBX3蛋白表达量(0.181±0.002)低于对照组(0.311±0.007)。PBX3过表达M320d组子宫内膜癌细胞中α-Catenin、E-cadherin蛋白表达量(0.139±0.019、0.143±0.007)低于对照组(0.399±0.034、0.261±0.017),Vimentin蛋白表达量(0.234±0.008)高于对照组(0.105±0.009),差异均有统计学意义(P<0.05)。双荧光素酶检验结果显示PBX3为miR-320d的下游靶基因。【结论】miR-320d可能通过降低下游靶基因PBX3水平影响EMT相关蛋白表达,抑制子宫内膜癌JEC细胞的上皮-间质转化功能。
