Aberrant RNA modification has been linked to the pathogenesis of various diseases;however,its specific molecular mechanisms in spinal cord injury remain poorly understood.The objective of this study was to explore RNA...Aberrant RNA modification has been linked to the pathogenesis of various diseases;however,its specific molecular mechanisms in spinal cord injury remain poorly understood.The objective of this study was to explore RNA modification-related biomarkers of spinal cord injury.The mRNA expression profiles of mice with spinal cord injury were retrieved from the Gene Expression Omnibus(GEO)database(GSE18179).We identified 185 differentially expressed genes using bioinformatics approaches.Functional enrichment analysis demonstrated aberrant activation or inhibition of common metabolism-related pathways,including sulfur metabolism and steroid biosynthesis,in mice with spinal cord injury.An integrated strategy comprising weighted gene co-expression network analysis,a random forest model,a support vector machine model,and a generalized linear model was employed to identify four genes whose aberrant RNA modification was linked to spinal cord injury:Elovl6,Idi1,Sqle,and Stbd1.We verified the expression levels and diagnostic performance of these four genes in the original training dataset and mouse samples via receiver operating characteristic curve analysis.Quantitative reverse transcription-polymerase chain reaction demonstrated variations in the mRNA levels of the four genes between the Sham and spinal cord injury groups at different time points following injury.We also constructed microRNA-mRNA and transcription factor-mRNA interaction networks using Cytoscape.Additionally,we evaluated the proportions of 22 types of immune cells in the spinal cords of mice using the CIBERSORT tool,revealing significant alterations in the numbers of memory B cells,resting dendritic cells,M0 macrophages,activated mast cells,resting mast cells,and CD8+T cells in spinal cord injury mice compared with Sham controls.Microglia and T cells were identified as key cell types by single-cell sequencing analysis.These findings provide new directions for the development of RNA modification-related therapeutic strategies for spinal cord injury and suggest that Elovl6,Idi1,Sqle,and Stbd1 are potential biomarkers of spinal cord injury.展开更多
目的研究胰岛素样生长因子2 mRNA结合蛋白1(IGF2BP1)及长链非编码RNA LINC00160(LINC00160)在胃癌中的作用,及其可能调控胃癌细胞增殖、侵袭的潜在作用机制。方法通过实时定量聚合酶链反应(qRT-PCR)检测胃癌组织和细胞中LINC00160表达...目的研究胰岛素样生长因子2 mRNA结合蛋白1(IGF2BP1)及长链非编码RNA LINC00160(LINC00160)在胃癌中的作用,及其可能调控胃癌细胞增殖、侵袭的潜在作用机制。方法通过实时定量聚合酶链反应(qRT-PCR)检测胃癌组织和细胞中LINC00160表达水平。通过生物信息学预测及RNA结合蛋白免疫沉淀(RIP)和甲基化RNA免疫沉淀(MeRIP)法验证分析LINC00160与IGF2BP1的结合作用;Pearson检验分析胃癌组织中LINC00160与IGF2BP1表达的相关性;CCK-8法和Transwell检测细胞增殖和侵袭能力;检测分析有氧糖酵解指标[葡萄糖摄取量、乳酸生成量和三磷酸腺苷(ATP)及细胞外酸化率(ECAR)和耗氧率(OCR)]水平变化。结果与癌旁正常组织相比,胃癌组织中LINC00160(5.13±0.62 vs 1.02±0.03)表达量显著上调,差异具有统计学意义(t=-36.266,P<0.001);胃癌细胞中LINC00160表达量亦高于人正常胃上皮细胞系GES-1,差异具有统计学意义(F=24.595,P<0.001)。与对照组相比,沉默LINC00160显著抑制AGS细胞的增殖(0.42±0.03 vs 1.03±0.04)和侵袭(22.13%±1.97%vs 42.15%±2.67%),减少葡萄糖摄取(2.11±0.26mmol/L vs 4.22±0.37mmol/L)和乳酸生成量(6.84±1.25mmol/L vs 11.68±1.55mmol/L),降低ECAR,升高ATP(3.34±0.29mmol/L vs 1.87±0.24mmol/L)水平和OCR,差异具有统计学意义(t=4.188~24.423,均P<0.01)。胃癌组织中IGF2BP1蛋白(4.07±0.36)表达显著高于癌旁组织(1.01±0.03),差异具有统计学意义(t=-46.396,P<0.01),且与LINC00160表达呈正相关(r 2=0.7745,P<0.01)。IGF2BP1通过与LINC00160的m6A修饰位点结合促进LINC00160稳定性。沉默IGF2BP1显著抑制了LINC00160表达及胃癌细胞的增殖、侵袭和有氧糖酵解,差异具有统计学意义(t=4.386~11.989,均P<0.01);过表达LINC00160可逆转沉默IGF2BP1对胃癌细胞生物学行为及有氧糖酵解的影响。结论胃癌中LINC00160显著上调,IGF2BP1可能通过m6A修饰稳定调控LINC00160表达,促进肿瘤细胞的有氧糖酵解,参与胃癌的发生发展。展开更多
文摘Aberrant RNA modification has been linked to the pathogenesis of various diseases;however,its specific molecular mechanisms in spinal cord injury remain poorly understood.The objective of this study was to explore RNA modification-related biomarkers of spinal cord injury.The mRNA expression profiles of mice with spinal cord injury were retrieved from the Gene Expression Omnibus(GEO)database(GSE18179).We identified 185 differentially expressed genes using bioinformatics approaches.Functional enrichment analysis demonstrated aberrant activation or inhibition of common metabolism-related pathways,including sulfur metabolism and steroid biosynthesis,in mice with spinal cord injury.An integrated strategy comprising weighted gene co-expression network analysis,a random forest model,a support vector machine model,and a generalized linear model was employed to identify four genes whose aberrant RNA modification was linked to spinal cord injury:Elovl6,Idi1,Sqle,and Stbd1.We verified the expression levels and diagnostic performance of these four genes in the original training dataset and mouse samples via receiver operating characteristic curve analysis.Quantitative reverse transcription-polymerase chain reaction demonstrated variations in the mRNA levels of the four genes between the Sham and spinal cord injury groups at different time points following injury.We also constructed microRNA-mRNA and transcription factor-mRNA interaction networks using Cytoscape.Additionally,we evaluated the proportions of 22 types of immune cells in the spinal cords of mice using the CIBERSORT tool,revealing significant alterations in the numbers of memory B cells,resting dendritic cells,M0 macrophages,activated mast cells,resting mast cells,and CD8+T cells in spinal cord injury mice compared with Sham controls.Microglia and T cells were identified as key cell types by single-cell sequencing analysis.These findings provide new directions for the development of RNA modification-related therapeutic strategies for spinal cord injury and suggest that Elovl6,Idi1,Sqle,and Stbd1 are potential biomarkers of spinal cord injury.
文摘目的研究胰岛素样生长因子2 mRNA结合蛋白1(IGF2BP1)及长链非编码RNA LINC00160(LINC00160)在胃癌中的作用,及其可能调控胃癌细胞增殖、侵袭的潜在作用机制。方法通过实时定量聚合酶链反应(qRT-PCR)检测胃癌组织和细胞中LINC00160表达水平。通过生物信息学预测及RNA结合蛋白免疫沉淀(RIP)和甲基化RNA免疫沉淀(MeRIP)法验证分析LINC00160与IGF2BP1的结合作用;Pearson检验分析胃癌组织中LINC00160与IGF2BP1表达的相关性;CCK-8法和Transwell检测细胞增殖和侵袭能力;检测分析有氧糖酵解指标[葡萄糖摄取量、乳酸生成量和三磷酸腺苷(ATP)及细胞外酸化率(ECAR)和耗氧率(OCR)]水平变化。结果与癌旁正常组织相比,胃癌组织中LINC00160(5.13±0.62 vs 1.02±0.03)表达量显著上调,差异具有统计学意义(t=-36.266,P<0.001);胃癌细胞中LINC00160表达量亦高于人正常胃上皮细胞系GES-1,差异具有统计学意义(F=24.595,P<0.001)。与对照组相比,沉默LINC00160显著抑制AGS细胞的增殖(0.42±0.03 vs 1.03±0.04)和侵袭(22.13%±1.97%vs 42.15%±2.67%),减少葡萄糖摄取(2.11±0.26mmol/L vs 4.22±0.37mmol/L)和乳酸生成量(6.84±1.25mmol/L vs 11.68±1.55mmol/L),降低ECAR,升高ATP(3.34±0.29mmol/L vs 1.87±0.24mmol/L)水平和OCR,差异具有统计学意义(t=4.188~24.423,均P<0.01)。胃癌组织中IGF2BP1蛋白(4.07±0.36)表达显著高于癌旁组织(1.01±0.03),差异具有统计学意义(t=-46.396,P<0.01),且与LINC00160表达呈正相关(r 2=0.7745,P<0.01)。IGF2BP1通过与LINC00160的m6A修饰位点结合促进LINC00160稳定性。沉默IGF2BP1显著抑制了LINC00160表达及胃癌细胞的增殖、侵袭和有氧糖酵解,差异具有统计学意义(t=4.386~11.989,均P<0.01);过表达LINC00160可逆转沉默IGF2BP1对胃癌细胞生物学行为及有氧糖酵解的影响。结论胃癌中LINC00160显著上调,IGF2BP1可能通过m6A修饰稳定调控LINC00160表达,促进肿瘤细胞的有氧糖酵解,参与胃癌的发生发展。
