目的 探析Xα1型胶原蛋白(collagen type X alpha 1,COL10A1)、微小RNA(microRNA,miRNA)-21、miRNA-320c表达水平对胃癌腹膜转移及预后的预测效能。方法 回顾性分析内蒙古医科大学附属肿瘤医院2023年7月至2024年9月收治的120例胃癌患者...目的 探析Xα1型胶原蛋白(collagen type X alpha 1,COL10A1)、微小RNA(microRNA,miRNA)-21、miRNA-320c表达水平对胃癌腹膜转移及预后的预测效能。方法 回顾性分析内蒙古医科大学附属肿瘤医院2023年7月至2024年9月收治的120例胃癌患者的临床资料。治疗后采用门诊及电话随访形式随访患者预后结局,并根据是否发生腹膜转移分为腹膜转移组和无腹膜转移组;根据预后结局分为不良组和良好组。采用酶联免疫法检测COL10A1;采用qRT-PCR检测miRNA-21、miRNA-320c。对比腹膜转移组、无腹膜转移组临床特征、血清COL10A1、miRNA-21、miRNA-320c;对比不同预后胃癌患者血清COL10A1、miRNA-21、miRNA-320c表达水平;采用Cox单因素及多因素对胃癌预后影响因素进行逐步回归分析;采用ROC分析血清COL10A1、miRNA-21、miRNA-320c对胃癌预后的预测效能。结果 120例胃癌患者中,腹膜转移31例,无腹膜转移89例。腹膜转移组TNM分期高于无腹膜转移组(P<0.05)。腹膜转移组血清COL10A1、miRNA-21、miRNA-320c均高于无腹膜转移组(P<0.05)。120例胃癌患者中,不良组42例,良好组78例。不良组血清COL10A1、miRNA-21、miRNA-320c均高于良好组(P<0.05)。Cox单因素显示,TNM分期、腹膜转移、淋巴结转移、COL10A1、miRNA-21、miRNA-320c与胃癌患者预后相关(均P<0.05);多因素显示,TNM分期、腹膜转移、COL10A1、miRNA-21、miRNA-320c是影响胃癌患者预后的独立危险因素(均P<0.05)。ROC曲线显示,血清COL10A1、miRNA-21、miRNA-320c联合预测胃癌预后结局的灵敏度及特异度分别为89.45%、92.34%,AUC达0.921,高于COL10A1(P=0.011)、miRNA-21(P=0.023)及miRNA-320c(P=0.015)单独诊断。结论 血清COL10A1、miRNA-21、miRNA-320c在胃癌腹膜转移及不良预后患者中呈高表达水平,且上述指标为胃癌患者不良预后危险因素,联合应用对预后结局有着较高预测效能。展开更多
本研究初步探讨过表达miRNA-320a对抑制神经胶质瘤U251细胞迁移、侵袭的可能的机制。实验开始前我们利用生物信息学软件进行分析对比miRNA-320a与水通道蛋白4(AQP4)之间的靶点结合关系,然后我们用miRNA-320a mimic及Ncontrol对U251细胞...本研究初步探讨过表达miRNA-320a对抑制神经胶质瘤U251细胞迁移、侵袭的可能的机制。实验开始前我们利用生物信息学软件进行分析对比miRNA-320a与水通道蛋白4(AQP4)之间的靶点结合关系,然后我们用miRNA-320a mimic及Ncontrol对U251细胞株进行转染,48 h后进行下一步实验。首先用q-PCR验证过表达转染情况,以及水通道蛋白4(AQP4)m RNA的表达水平,其次用划痕和Transwell检测转染后细胞株的迁移侵袭能力,最后用Western blotting测定AQP4的表达水平。生物信息分析可得miRNA-320a在AQP4 m RNA 3'UTR区域能稳定结合,实验结果显示转染mimic后,过表达组明显升高,且过表达组AQP4 m RNA的表达明显被抑制,划痕和Transwell实验提示了过表达miRNA-320a后能抑制U251细胞株的迁移侵袭能力(p<0.01)。Western blotting结果显示,过表达miRNA-320a与对照组相比能明显抑制AQP4蛋白的表达。所有研究结果提示miRNA-320a能靶向作用AQP4 m RNA 3'UTR区域,并抑制其蛋白表达,从而抑制了肿瘤细胞U251的迁移侵袭能力,为临床治疗恶性胶质瘤提供新的参考。展开更多
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
BACKGROUND Exosomal miRNAs play crucial roles in many central nervous system diseases.Cerebral small vessel disease(CVSD)is a small vessel disease that is affected by various factors.This study aimed to investigate th...BACKGROUND Exosomal miRNAs play crucial roles in many central nervous system diseases.Cerebral small vessel disease(CVSD)is a small vessel disease that is affected by various factors.This study aimed to investigate the role of exosomal miR-320e in the Wnt/β-catenin pathway stimulated by oxidative stress and assess its clinical correlation with psychiatric symptoms in patients with CVSD.AIM To explore whether exosomal miR-320e could suppress the Wnt/β-catenin pathway and play a protective role in CVSD progression,as well as examine its potential correlation with cognitive impairment and depression in patients with CVSD.METHODS Differentially expressed exosomal miRNAs were filtered by sequencing plasma exosomes from patients with CVSD and healthy controls.Bioinformatics and dual luciferase analyses were used to confirm the binding of miR-320e to Wnt2,and the mRNA and protein levels of downstream components in the Wnt/β-catenin pathway were evaluated when overexpressed or with knockdown of miR-320e under H2O2-induced oxidative stress.In addition,Wnt2-targeting siRNA was used to confirm the role of miR-320e in the Wnt2-mediated inhibition of the Wnt/β-catenin pathway.A retrospective analysis was conducted among patients with CVSD to confirm the correlation between miR-320e expression and the severity of cognitive impairment and depression,which were quantified using the Montreal Cognitive Assessment(MoCA)/Executive Function Assessment(EFA),and the Hamilton Depression Scale(HAMD)/Beck Depression Inventory(BDI),respectively.RESULTS High-throughput sequencing revealed that exosomal miR-320e was downregulated in patients with CVSD.Bioinformatics analysis and dual-luciferase reporter gene experiments showed that exosomal miR-320e inhibited the Wnt/β-catenin pathway in response to oxidative stress by targeting the 3'noncoding region of Wnt2.Uptake of exosomes carrying miR-320e into endothelial cells could also target Wnt2 and inhibit the Wnt2/β-catenin pathway.Elevated miR-320e expression may protect patients with CVSD from relatively severe cognitive impairment and depression,as it was found to have a positive correlation with the MoCA/EFA and HAMD/BDI scores.CONCLUSION Our results suggest that exosomal miR-320e suppresses the Wnt/β-catenin pathway and may play a protective role in CVSD progression.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.
文摘目的 探析Xα1型胶原蛋白(collagen type X alpha 1,COL10A1)、微小RNA(microRNA,miRNA)-21、miRNA-320c表达水平对胃癌腹膜转移及预后的预测效能。方法 回顾性分析内蒙古医科大学附属肿瘤医院2023年7月至2024年9月收治的120例胃癌患者的临床资料。治疗后采用门诊及电话随访形式随访患者预后结局,并根据是否发生腹膜转移分为腹膜转移组和无腹膜转移组;根据预后结局分为不良组和良好组。采用酶联免疫法检测COL10A1;采用qRT-PCR检测miRNA-21、miRNA-320c。对比腹膜转移组、无腹膜转移组临床特征、血清COL10A1、miRNA-21、miRNA-320c;对比不同预后胃癌患者血清COL10A1、miRNA-21、miRNA-320c表达水平;采用Cox单因素及多因素对胃癌预后影响因素进行逐步回归分析;采用ROC分析血清COL10A1、miRNA-21、miRNA-320c对胃癌预后的预测效能。结果 120例胃癌患者中,腹膜转移31例,无腹膜转移89例。腹膜转移组TNM分期高于无腹膜转移组(P<0.05)。腹膜转移组血清COL10A1、miRNA-21、miRNA-320c均高于无腹膜转移组(P<0.05)。120例胃癌患者中,不良组42例,良好组78例。不良组血清COL10A1、miRNA-21、miRNA-320c均高于良好组(P<0.05)。Cox单因素显示,TNM分期、腹膜转移、淋巴结转移、COL10A1、miRNA-21、miRNA-320c与胃癌患者预后相关(均P<0.05);多因素显示,TNM分期、腹膜转移、COL10A1、miRNA-21、miRNA-320c是影响胃癌患者预后的独立危险因素(均P<0.05)。ROC曲线显示,血清COL10A1、miRNA-21、miRNA-320c联合预测胃癌预后结局的灵敏度及特异度分别为89.45%、92.34%,AUC达0.921,高于COL10A1(P=0.011)、miRNA-21(P=0.023)及miRNA-320c(P=0.015)单独诊断。结论 血清COL10A1、miRNA-21、miRNA-320c在胃癌腹膜转移及不良预后患者中呈高表达水平,且上述指标为胃癌患者不良预后危险因素,联合应用对预后结局有着较高预测效能。
文摘本研究初步探讨过表达miRNA-320a对抑制神经胶质瘤U251细胞迁移、侵袭的可能的机制。实验开始前我们利用生物信息学软件进行分析对比miRNA-320a与水通道蛋白4(AQP4)之间的靶点结合关系,然后我们用miRNA-320a mimic及Ncontrol对U251细胞株进行转染,48 h后进行下一步实验。首先用q-PCR验证过表达转染情况,以及水通道蛋白4(AQP4)m RNA的表达水平,其次用划痕和Transwell检测转染后细胞株的迁移侵袭能力,最后用Western blotting测定AQP4的表达水平。生物信息分析可得miRNA-320a在AQP4 m RNA 3'UTR区域能稳定结合,实验结果显示转染mimic后,过表达组明显升高,且过表达组AQP4 m RNA的表达明显被抑制,划痕和Transwell实验提示了过表达miRNA-320a后能抑制U251细胞株的迁移侵袭能力(p<0.01)。Western blotting结果显示,过表达miRNA-320a与对照组相比能明显抑制AQP4蛋白的表达。所有研究结果提示miRNA-320a能靶向作用AQP4 m RNA 3'UTR区域,并抑制其蛋白表达,从而抑制了肿瘤细胞U251的迁移侵袭能力,为临床治疗恶性胶质瘤提供新的参考。
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BACKGROUND Exosomal miRNAs play crucial roles in many central nervous system diseases.Cerebral small vessel disease(CVSD)is a small vessel disease that is affected by various factors.This study aimed to investigate the role of exosomal miR-320e in the Wnt/β-catenin pathway stimulated by oxidative stress and assess its clinical correlation with psychiatric symptoms in patients with CVSD.AIM To explore whether exosomal miR-320e could suppress the Wnt/β-catenin pathway and play a protective role in CVSD progression,as well as examine its potential correlation with cognitive impairment and depression in patients with CVSD.METHODS Differentially expressed exosomal miRNAs were filtered by sequencing plasma exosomes from patients with CVSD and healthy controls.Bioinformatics and dual luciferase analyses were used to confirm the binding of miR-320e to Wnt2,and the mRNA and protein levels of downstream components in the Wnt/β-catenin pathway were evaluated when overexpressed or with knockdown of miR-320e under H2O2-induced oxidative stress.In addition,Wnt2-targeting siRNA was used to confirm the role of miR-320e in the Wnt2-mediated inhibition of the Wnt/β-catenin pathway.A retrospective analysis was conducted among patients with CVSD to confirm the correlation between miR-320e expression and the severity of cognitive impairment and depression,which were quantified using the Montreal Cognitive Assessment(MoCA)/Executive Function Assessment(EFA),and the Hamilton Depression Scale(HAMD)/Beck Depression Inventory(BDI),respectively.RESULTS High-throughput sequencing revealed that exosomal miR-320e was downregulated in patients with CVSD.Bioinformatics analysis and dual-luciferase reporter gene experiments showed that exosomal miR-320e inhibited the Wnt/β-catenin pathway in response to oxidative stress by targeting the 3'noncoding region of Wnt2.Uptake of exosomes carrying miR-320e into endothelial cells could also target Wnt2 and inhibit the Wnt2/β-catenin pathway.Elevated miR-320e expression may protect patients with CVSD from relatively severe cognitive impairment and depression,as it was found to have a positive correlation with the MoCA/EFA and HAMD/BDI scores.CONCLUSION Our results suggest that exosomal miR-320e suppresses the Wnt/β-catenin pathway and may play a protective role in CVSD progression.
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.
