目的探讨miR-590-5p、DNA损伤检查点蛋白调节子1(mediator of DNA damage checkpoint 1,MDC1)在高级别胶质瘤(high-grade glioma,HGG)组织中的表达及与胶质瘤病人术后放疗效果的关系,并明确二者对胶质瘤细胞增殖、凋亡的影响。方法选取2...目的探讨miR-590-5p、DNA损伤检查点蛋白调节子1(mediator of DNA damage checkpoint 1,MDC1)在高级别胶质瘤(high-grade glioma,HGG)组织中的表达及与胶质瘤病人术后放疗效果的关系,并明确二者对胶质瘤细胞增殖、凋亡的影响。方法选取2019年1月至2021年2月河北北方学院附属第一医院64例HGG患者,评估放疗效果。实时荧光定量PCR(qRT-PCR)法检测miR-590-5p水平,免疫组织化学染色检测MDC1表达情况,分析miR-590-5p、MDC1表达与胶质瘤病人术后放疗效果的关系,多因素Logistic回归分析影响HGG患者术后放疗效果的因素;体外培养胶质瘤U87MG细胞,并分别转染miR-590-5p mimic、MDC1-shRNA及其阴性对照,CCK-8法和流式细胞术分别检测细胞增殖和凋亡;构建裸鼠移植瘤模型,观察过表达miR-590-5p和敲低MDC1对肿瘤生长的影响。结果MDC1在HGG组织中的表达较正常脑组织中升高,mi R-590-5p表达较正常脑组织降低,二者表达水平呈负相关;MDC1表达升高、miR-590-5p表达降低,其放疗效果越差;Logistic回归分析显示,MDC1高表达、miR-590-5p低表达均是影响HGG患者放疗效果的危险因素。过表达miR-590-5p和敲低MDC1后,U87MG细胞增殖力降低,凋亡率升高,移植瘤体积和重量下降,Ki-67阳性细胞比例减少。过表达miR-590-5p后MDC1蛋白表达明显下降。结论HGG组织中miR-590-5p呈低表达,MDC1呈高表达,二者表达与HGG的发生和患者术后放疗效果关系密切;过表达miR-590-5p和敲低MDC1表达可抑制胶质瘤细胞增殖并促进凋亡。展开更多
BACKGROUND As reported,multiple circular RNAs(circRNAs)interfere with colorectal cancer(CRC)progression.Here,circRNA_0001658(circ_0001658)is focused on studying how it works in CRC.AIM Clarify the expression pattern,b...BACKGROUND As reported,multiple circular RNAs(circRNAs)interfere with colorectal cancer(CRC)progression.Here,circRNA_0001658(circ_0001658)is focused on studying how it works in CRC.AIM Clarify the expression pattern,biological function,and underlying mechanism of circ_0001658 of CRC tumorigenesis.METHODS In CRC-related chip data retrieved using the database named Gene Expression Omnibus,different expressions of circRNAs between CRC and normal tissue samples were identified.Quantitative Real-time PCR and Western blot ensured the analysis on circ_0001658,microRNA-590-5P(miR-590-5p),and methyltransferase-like 3(METTL3)mRNA expressions in tissues and cells.Cell counting kit-8 and flow cytometry were used to detect cell proliferation,apoptosis and migration.The targeting relations between circ_0001658,miR-590-5p,and METTL3 mRNA 3'-untranslated region were under the verification of bioinformatics prediction and dual luciferase-based reporter gene assays.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis were employed on the downstream targets of miR-590-5p using the Database for Annotation,Visualization and Integrated Discovery database.RESULTS Circ_0001658 and METTL3 mRNA was elevated in CRC tissues and cells,whereas miR-590-5p was decreased.Circ_0001658 overexpression promoted the proliferation of HT29 cells,inhibited apoptosis,and accelerated the cell cycle.In SW480 cells,knocking down circ_0001658 had the opposite effect.Circ_0001658 could specifically bind to miR-590-5p and negatively modulate its expressions;METTL3 is a miR-590-5p target that can be positively regulated by circ 0001658.Circ 0001658 was inversely associated with miR-590-5p expression while positively with METTL3 expressions.CONCLUSION Circ_0001658 regulates the miR-590-5p/METTL 3-axis to increase CRC cell growth and decrease apoptosis.展开更多
文摘目的探讨miR-590-5p、DNA损伤检查点蛋白调节子1(mediator of DNA damage checkpoint 1,MDC1)在高级别胶质瘤(high-grade glioma,HGG)组织中的表达及与胶质瘤病人术后放疗效果的关系,并明确二者对胶质瘤细胞增殖、凋亡的影响。方法选取2019年1月至2021年2月河北北方学院附属第一医院64例HGG患者,评估放疗效果。实时荧光定量PCR(qRT-PCR)法检测miR-590-5p水平,免疫组织化学染色检测MDC1表达情况,分析miR-590-5p、MDC1表达与胶质瘤病人术后放疗效果的关系,多因素Logistic回归分析影响HGG患者术后放疗效果的因素;体外培养胶质瘤U87MG细胞,并分别转染miR-590-5p mimic、MDC1-shRNA及其阴性对照,CCK-8法和流式细胞术分别检测细胞增殖和凋亡;构建裸鼠移植瘤模型,观察过表达miR-590-5p和敲低MDC1对肿瘤生长的影响。结果MDC1在HGG组织中的表达较正常脑组织中升高,mi R-590-5p表达较正常脑组织降低,二者表达水平呈负相关;MDC1表达升高、miR-590-5p表达降低,其放疗效果越差;Logistic回归分析显示,MDC1高表达、miR-590-5p低表达均是影响HGG患者放疗效果的危险因素。过表达miR-590-5p和敲低MDC1后,U87MG细胞增殖力降低,凋亡率升高,移植瘤体积和重量下降,Ki-67阳性细胞比例减少。过表达miR-590-5p后MDC1蛋白表达明显下降。结论HGG组织中miR-590-5p呈低表达,MDC1呈高表达,二者表达与HGG的发生和患者术后放疗效果关系密切;过表达miR-590-5p和敲低MDC1表达可抑制胶质瘤细胞增殖并促进凋亡。
文摘BACKGROUND As reported,multiple circular RNAs(circRNAs)interfere with colorectal cancer(CRC)progression.Here,circRNA_0001658(circ_0001658)is focused on studying how it works in CRC.AIM Clarify the expression pattern,biological function,and underlying mechanism of circ_0001658 of CRC tumorigenesis.METHODS In CRC-related chip data retrieved using the database named Gene Expression Omnibus,different expressions of circRNAs between CRC and normal tissue samples were identified.Quantitative Real-time PCR and Western blot ensured the analysis on circ_0001658,microRNA-590-5P(miR-590-5p),and methyltransferase-like 3(METTL3)mRNA expressions in tissues and cells.Cell counting kit-8 and flow cytometry were used to detect cell proliferation,apoptosis and migration.The targeting relations between circ_0001658,miR-590-5p,and METTL3 mRNA 3'-untranslated region were under the verification of bioinformatics prediction and dual luciferase-based reporter gene assays.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis were employed on the downstream targets of miR-590-5p using the Database for Annotation,Visualization and Integrated Discovery database.RESULTS Circ_0001658 and METTL3 mRNA was elevated in CRC tissues and cells,whereas miR-590-5p was decreased.Circ_0001658 overexpression promoted the proliferation of HT29 cells,inhibited apoptosis,and accelerated the cell cycle.In SW480 cells,knocking down circ_0001658 had the opposite effect.Circ_0001658 could specifically bind to miR-590-5p and negatively modulate its expressions;METTL3 is a miR-590-5p target that can be positively regulated by circ 0001658.Circ 0001658 was inversely associated with miR-590-5p expression while positively with METTL3 expressions.CONCLUSION Circ_0001658 regulates the miR-590-5p/METTL 3-axis to increase CRC cell growth and decrease apoptosis.
