Objective This study aimed to examine the role of long non-coding RNA PCED1B antisense RNA 1(PCED1B-AS1)in the development of hepatocellular carcinoma(HCC).Methods A total of 62 pairs of HCC tissues and adjacent non-t...Objective This study aimed to examine the role of long non-coding RNA PCED1B antisense RNA 1(PCED1B-AS1)in the development of hepatocellular carcinoma(HCC).Methods A total of 62 pairs of HCC tissues and adjacent non-tumor tissues were obtained from 62 HCC patients.The interactions of PCED1B-AS1 and microRNA-34a(miR-34a)were detected by dual luciferase activity assay and RNA pull-down assay.The RNA expression levels of PCED1B-AS1,miR-34a and CD44 were detected by RT-qPCR,and the protein expression level of CD44 was determined by Western blotting.The cell proliferation was detected by cell proliferation assay,and the cell invasion and migration by transwell invasion assay.The HCC tumor growth after PCED1B-AS1 was downregulated was determined by in vivo animal study.Results PCED1B-AS1 was highly expressed in HCC tissues,which was associated with poor survival of HCC patients.Furthermore,PCED1B-AS1 interacted with miR-34a in HCC cells,but they did not regulate the expression of each other.Additionally,PCED1B-AS1 increased the expression level of CD44,which was targeted by miR-34a.The cell proliferation and invasion assay revealed that miR-34a inhibited the proliferation and invasion of HCC in vitro,while CD44 exhibited the opposite effects.Furthermore,PCED1B-AS1 suppressed the role of miR-34a.Moreover,the knockdown of PCED1B-AS1 repressed the HCC tumor growth in nude mice in vivo.Conclusion PCED1B-AS1 may play an oncogenic role by regulating the miR-34a/CD44 axis in HCC.展开更多
Background:Aortic dissection(AD)is a fatal cardiovascular disease for which the key involved genes are largely unknown.Here,we aimed to identify promising AD biomarkers from high-throughput RNA expressing data.Methods...Background:Aortic dissection(AD)is a fatal cardiovascular disease for which the key involved genes are largely unknown.Here,we aimed to identify promising AD biomarkers from high-throughput RNA expressing data.Methods:In the GSE98770 dataset,differentially expressed mRNAs(DE-mRNAs)and microRNAs(DE-microRNAs)were identified through differentially expressed gene analysis and gene set enrichment analysis.The regulatory network between DE-mRNAs and DE-microRNAs was established,and hub genes were identified with Cytoscape.Relationships between hub genes and AD were confirmed in the Comparative Toxicogenomics Database(CTD).Potential key transcription factors were discovered with Cytoscape.Hub gene verification was performed by qPCR and immunofluorescence analyses of human specimens.Results:DE-mRNAs and DE-microRNAs were identified.Four mRNAs and microRNA-1321(miR-1321)were found to have the most connections with other genes.CBL was connected to the most genes and interacted with miR-1321,which was also connected to the most genes among the DE-microRNAs.In addition,CBL was associated with AD in the CTD.Among the top five transcription factors potentially regulating CBL transcription,only HOXB13 was a DE-mRNA.The findings were further successfully verified in human specimens.Conclusion:CBL,which may be transcriptionally regulated by HOXB13 and post-transcriptionally regulated by miR-1321,was identified as the most promising potential biomarker for AD.展开更多
Acute mitochondrial damage and the energy crisis following axonal injury highlight mitochondrial transport as an important target for axonal regeneration.Syntaphilin(Snph),known for its potent mitochondrial anchoring ...Acute mitochondrial damage and the energy crisis following axonal injury highlight mitochondrial transport as an important target for axonal regeneration.Syntaphilin(Snph),known for its potent mitochondrial anchoring action,has emerged as a significant inhibitor of both mitochondrial transport and axonal regeneration.Therefore,investigating the molecular mechanisms that influence the expression levels of the snph gene can provide a viable strategy to regulate mitochondrial trafficking and enhance axonal regeneration.Here,we reveal the inhibitory effect of microRNA-146b(miR-146b)on the expression of the homologous zebrafish gene syntaphilin b(snphb).Through CRISPR/Cas9 and single-cell electroporation,we elucidated the positive regulatory effect of the miR-146b-snphb axis on Mauthner cell(M-cell)axon regeneration at the global and single-cell levels.Through escape response tests,we show that miR-146b-snphb signaling positively regulates functional recovery after M-cell axon injury.In addition,continuous dynamic imaging in vivo showed that reprogramming miR-146b significantly promotes axonal mitochondrial trafficking in the pre-injury and early stages of regeneration.Our study reveals an intrinsic axonal regeneration regulatory axis that promotes axonal regeneration by reprogramming mitochondrial transport and anchoring.This regulation involves noncoding RNA,and mitochondria-associated genes may provide a potential opportunity for the repair of central nervous system injury.展开更多
BACKGROUND As a non-coding RNA molecule,circular RNAs(circRNAs)have significant specificity,and existing data suggest a close relationship between them and the prognosis of patients with gastric cancer(GC).However,thi...BACKGROUND As a non-coding RNA molecule,circular RNAs(circRNAs)have significant specificity,and existing data suggest a close relationship between them and the prognosis of patients with gastric cancer(GC).However,this mechanism has no evidence yet.This article explores the functions of hsa_circRNA_102415 in the malignant behavior and potential downstream signaling of GC cells.The chosen approach is loss of signal and functional gain.AIM To investigate and analyze the relationship between hsa_circRNA_102415 and GC and explore its specific role.Results provide reference for other researchers to develop targeted treatment plans.METHODS The gene expression omnibus(GEO)database can be used to obtain the microarray dataset GSE83521.Data were analyzed using the GEO2R tool to identify differences in circRNAs between normal and GC samples.Quantitative real-time polymerase chain reaction was used to detect differentially expressed genes in GC tissue samples and adjacent cancer tissue samples.GC cells were transfected with small interfering-hsa_circRNA_104415 and plasmid DNA(pcDNA)-hsa_ircRNA_102415.Multiple detection methods,such as Transwell and cell counting kit 8,were used to evaluate cellular physiological activities,including cell invasion and proliferation.The relationship between Wnt family members 2B,microRNA(miR)-4529-5p,etc.,including argonaute 2-RNA immunoprecipitation and luciferase reporter genes was analyzed.Rescue experiments were conducted to analyze and explore the relationship between the malignant behavior of GC cells and hsa_circRNA_102415.RESULTS GEO2R analysis confirmed that hsa_circRNA_102415 had significantly higher expression levels in disease tissues.hsa_circRNA_102415 and miR-4529-5p showed a negative correlation in disease cells,suggesting that hsa_circRNA_102415 upregulated WNT2B expression in GC cells as a competing endogenous RNA for miR-4529-5p.miR-4529-5p mimic or small interfering-WNT2B reversed the effects of pcDNA-hsa_circRNA_102415 or miR-4529-5p inhibitor on cell malignant functions.CONCLUSION miR-4529-5p was used to successfully activate the potential of WNT2B,clarify the role of hsa_circRNA_102415 in GC cells,and provide reference for other researchers to develop targeted treatment plans.展开更多
目的研究miR-21在弥漫大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)中的表达,探讨miR-21表达与DL-BCL临床病理特征的关系及其在DLBCL发生发展中的意义。方法采用Real-time RT-PCR方法检测36例DLBCL和10例正常淋巴结中miR-21的表...目的研究miR-21在弥漫大B细胞淋巴瘤(diffuse large B-cell lymphoma,DLBCL)中的表达,探讨miR-21表达与DL-BCL临床病理特征的关系及其在DLBCL发生发展中的意义。方法采用Real-time RT-PCR方法检测36例DLBCL和10例正常淋巴结中miR-21的表达,并采用免疫组织化学SP法检测Ki-67、PTEN在DLBCL中的表达。结果miR-21在DLBCL中高表达,36例DLBCL中有14例表达PTEN(38.9%),21例Ki-67≥50%(58.3%)。DLBCL中miR-21表达水平与PTEN蛋白表达呈负相关,其高表达与DLBCL高Ann Arbor分期、高增殖指数(Ki-67≥50%)、国际预后指数(IPI)呈正相关。结论miR-21过表达可能是DLBCL恶性度高的标志,是促进DLBCL肿瘤细胞增殖的重要因素。PTEN可能是miR-21在DLBCL发挥作用的靶标。展开更多
目的探讨催产素(oxytocin,OXT)对卵巢癌细胞增殖与凋亡的影响及其作用机制。方法人卵巢癌细胞系SKOV3和A2780细胞来源于美国模式培养物保藏所(ATCC)。OXT和OXT受体(oxytocin receptor,OXTR)拮抗剂Atosiban刺激卵巢癌细胞,改变细胞中miR-...目的探讨催产素(oxytocin,OXT)对卵巢癌细胞增殖与凋亡的影响及其作用机制。方法人卵巢癌细胞系SKOV3和A2780细胞来源于美国模式培养物保藏所(ATCC)。OXT和OXT受体(oxytocin receptor,OXTR)拮抗剂Atosiban刺激卵巢癌细胞,改变细胞中miR-196b-3p的水平,使用MTT法、ELISA试验、Western blot、生物信息学工具及荧光素酶报告基因实验等研究以上处理对卵巢癌细胞增殖与凋亡的作用和机制。结果OXT刺激SKOV3细胞OXTR激活(F=28.842,P<0.05),增殖活性降低(F=12.988,P<0.05),Caspase3活性增加(F=26.676,P<0.05);Ki67、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)及p53凋亡刺激蛋白抑制剂(inhibitor of apoptosis-stimulating of p53 protein,iASPP)表达下调,p53凋亡刺激蛋白(apoptosis-stimulating of p53 protein,ASPP)1(ASPP1)及2(ASPP2)表达上调;但Atosiban部分逆转OXT的效果。OXT刺激卵巢癌细胞miR-196b-3p水平减少(A2780:F=76.406,P<0.05;SKOV3:F=45.874,P<0.05)。miR-196b-3p-mimic转染联合OXT刺激,导致细胞增殖活性增加(F=9.232,P<0.05),Caspase3活性降低(F=36.350,P<0.05),同时ASPP2表达降低(F=83.013,P<0.05)。而miR-196b-3p-inhibitor转染联合OXT刺激,ASPP2表达增加(F=83.013,P<0.05)。生物信息学工具预测联合荧光素酶报告基因实验确认ASPP2是miR-196b-3p的靶基因。结论OXT介导的OXTR激活通过调节miR-196b-3p/ASPP2信号通路调节卵巢癌细胞生长。展开更多
基金supported by the Medical Science and Technology Research Foundation of Guangdong Province(No.A2020559).
文摘Objective This study aimed to examine the role of long non-coding RNA PCED1B antisense RNA 1(PCED1B-AS1)in the development of hepatocellular carcinoma(HCC).Methods A total of 62 pairs of HCC tissues and adjacent non-tumor tissues were obtained from 62 HCC patients.The interactions of PCED1B-AS1 and microRNA-34a(miR-34a)were detected by dual luciferase activity assay and RNA pull-down assay.The RNA expression levels of PCED1B-AS1,miR-34a and CD44 were detected by RT-qPCR,and the protein expression level of CD44 was determined by Western blotting.The cell proliferation was detected by cell proliferation assay,and the cell invasion and migration by transwell invasion assay.The HCC tumor growth after PCED1B-AS1 was downregulated was determined by in vivo animal study.Results PCED1B-AS1 was highly expressed in HCC tissues,which was associated with poor survival of HCC patients.Furthermore,PCED1B-AS1 interacted with miR-34a in HCC cells,but they did not regulate the expression of each other.Additionally,PCED1B-AS1 increased the expression level of CD44,which was targeted by miR-34a.The cell proliferation and invasion assay revealed that miR-34a inhibited the proliferation and invasion of HCC in vitro,while CD44 exhibited the opposite effects.Furthermore,PCED1B-AS1 suppressed the role of miR-34a.Moreover,the knockdown of PCED1B-AS1 repressed the HCC tumor growth in nude mice in vivo.Conclusion PCED1B-AS1 may play an oncogenic role by regulating the miR-34a/CD44 axis in HCC.
基金supported by grants from the Guangzhou Basic and Applied Basic Research Foundation(202201010940)National Natural Science Foundation of China(82200306)Guangdong Basic and Applied Basic Research Foundation(2021A1515111092).
文摘Background:Aortic dissection(AD)is a fatal cardiovascular disease for which the key involved genes are largely unknown.Here,we aimed to identify promising AD biomarkers from high-throughput RNA expressing data.Methods:In the GSE98770 dataset,differentially expressed mRNAs(DE-mRNAs)and microRNAs(DE-microRNAs)were identified through differentially expressed gene analysis and gene set enrichment analysis.The regulatory network between DE-mRNAs and DE-microRNAs was established,and hub genes were identified with Cytoscape.Relationships between hub genes and AD were confirmed in the Comparative Toxicogenomics Database(CTD).Potential key transcription factors were discovered with Cytoscape.Hub gene verification was performed by qPCR and immunofluorescence analyses of human specimens.Results:DE-mRNAs and DE-microRNAs were identified.Four mRNAs and microRNA-1321(miR-1321)were found to have the most connections with other genes.CBL was connected to the most genes and interacted with miR-1321,which was also connected to the most genes among the DE-microRNAs.In addition,CBL was associated with AD in the CTD.Among the top five transcription factors potentially regulating CBL transcription,only HOXB13 was a DE-mRNA.The findings were further successfully verified in human specimens.Conclusion:CBL,which may be transcriptionally regulated by HOXB13 and post-transcriptionally regulated by miR-1321,was identified as the most promising potential biomarker for AD.
基金supported by the core facility Center for Life Sciences,University of Science and Technology of China,Research Funds of the Center for Advanced Interdisciplinary Science and Biomedicine of IHM(QYZD20220002)the National Natural Science Foundation of China(82071357)the Ministry of Science and Technology of China(2019YFA0405600).
文摘Acute mitochondrial damage and the energy crisis following axonal injury highlight mitochondrial transport as an important target for axonal regeneration.Syntaphilin(Snph),known for its potent mitochondrial anchoring action,has emerged as a significant inhibitor of both mitochondrial transport and axonal regeneration.Therefore,investigating the molecular mechanisms that influence the expression levels of the snph gene can provide a viable strategy to regulate mitochondrial trafficking and enhance axonal regeneration.Here,we reveal the inhibitory effect of microRNA-146b(miR-146b)on the expression of the homologous zebrafish gene syntaphilin b(snphb).Through CRISPR/Cas9 and single-cell electroporation,we elucidated the positive regulatory effect of the miR-146b-snphb axis on Mauthner cell(M-cell)axon regeneration at the global and single-cell levels.Through escape response tests,we show that miR-146b-snphb signaling positively regulates functional recovery after M-cell axon injury.In addition,continuous dynamic imaging in vivo showed that reprogramming miR-146b significantly promotes axonal mitochondrial trafficking in the pre-injury and early stages of regeneration.Our study reveals an intrinsic axonal regeneration regulatory axis that promotes axonal regeneration by reprogramming mitochondrial transport and anchoring.This regulation involves noncoding RNA,and mitochondria-associated genes may provide a potential opportunity for the repair of central nervous system injury.
文摘BACKGROUND As a non-coding RNA molecule,circular RNAs(circRNAs)have significant specificity,and existing data suggest a close relationship between them and the prognosis of patients with gastric cancer(GC).However,this mechanism has no evidence yet.This article explores the functions of hsa_circRNA_102415 in the malignant behavior and potential downstream signaling of GC cells.The chosen approach is loss of signal and functional gain.AIM To investigate and analyze the relationship between hsa_circRNA_102415 and GC and explore its specific role.Results provide reference for other researchers to develop targeted treatment plans.METHODS The gene expression omnibus(GEO)database can be used to obtain the microarray dataset GSE83521.Data were analyzed using the GEO2R tool to identify differences in circRNAs between normal and GC samples.Quantitative real-time polymerase chain reaction was used to detect differentially expressed genes in GC tissue samples and adjacent cancer tissue samples.GC cells were transfected with small interfering-hsa_circRNA_104415 and plasmid DNA(pcDNA)-hsa_ircRNA_102415.Multiple detection methods,such as Transwell and cell counting kit 8,were used to evaluate cellular physiological activities,including cell invasion and proliferation.The relationship between Wnt family members 2B,microRNA(miR)-4529-5p,etc.,including argonaute 2-RNA immunoprecipitation and luciferase reporter genes was analyzed.Rescue experiments were conducted to analyze and explore the relationship between the malignant behavior of GC cells and hsa_circRNA_102415.RESULTS GEO2R analysis confirmed that hsa_circRNA_102415 had significantly higher expression levels in disease tissues.hsa_circRNA_102415 and miR-4529-5p showed a negative correlation in disease cells,suggesting that hsa_circRNA_102415 upregulated WNT2B expression in GC cells as a competing endogenous RNA for miR-4529-5p.miR-4529-5p mimic or small interfering-WNT2B reversed the effects of pcDNA-hsa_circRNA_102415 or miR-4529-5p inhibitor on cell malignant functions.CONCLUSION miR-4529-5p was used to successfully activate the potential of WNT2B,clarify the role of hsa_circRNA_102415 in GC cells,and provide reference for other researchers to develop targeted treatment plans.
文摘目的探讨催产素(oxytocin,OXT)对卵巢癌细胞增殖与凋亡的影响及其作用机制。方法人卵巢癌细胞系SKOV3和A2780细胞来源于美国模式培养物保藏所(ATCC)。OXT和OXT受体(oxytocin receptor,OXTR)拮抗剂Atosiban刺激卵巢癌细胞,改变细胞中miR-196b-3p的水平,使用MTT法、ELISA试验、Western blot、生物信息学工具及荧光素酶报告基因实验等研究以上处理对卵巢癌细胞增殖与凋亡的作用和机制。结果OXT刺激SKOV3细胞OXTR激活(F=28.842,P<0.05),增殖活性降低(F=12.988,P<0.05),Caspase3活性增加(F=26.676,P<0.05);Ki67、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)及p53凋亡刺激蛋白抑制剂(inhibitor of apoptosis-stimulating of p53 protein,iASPP)表达下调,p53凋亡刺激蛋白(apoptosis-stimulating of p53 protein,ASPP)1(ASPP1)及2(ASPP2)表达上调;但Atosiban部分逆转OXT的效果。OXT刺激卵巢癌细胞miR-196b-3p水平减少(A2780:F=76.406,P<0.05;SKOV3:F=45.874,P<0.05)。miR-196b-3p-mimic转染联合OXT刺激,导致细胞增殖活性增加(F=9.232,P<0.05),Caspase3活性降低(F=36.350,P<0.05),同时ASPP2表达降低(F=83.013,P<0.05)。而miR-196b-3p-inhibitor转染联合OXT刺激,ASPP2表达增加(F=83.013,P<0.05)。生物信息学工具预测联合荧光素酶报告基因实验确认ASPP2是miR-196b-3p的靶基因。结论OXT介导的OXTR激活通过调节miR-196b-3p/ASPP2信号通路调节卵巢癌细胞生长。
