RESUMO
Hsa_circ_0061395(circBACH1) and SERBP1(SERPINE1 mRNA binding protein 1) have been reported to play a carcinogenic role in HCC.In this study, circBACH1, microRNA(miR)-656-3p, and SERBP1 expression levels with quantitative real-time polymerase chain reaction (qRT-PCR) in HCC tissue specimens and cells.The protein levels of SERBP1, E-Cadherin, vimentin, and N-Cadherin were detected with western blotting.Cell proliferation, migration, invasion, and apoptosis were determined with CCK-8, colony formation, transwell, and flow cytometry assays.The targeting relatio-nship between circBACH1 or SERBP1 and miR-656-3p was verified by dual-lucifer- ase reporter assay.The role of circBACH1 was validated by xenograft assay.CircBAC- H1 and SERBP1 were upregulated in HCC tissues and cells.Both circBACH1 and SERBP1 knockdown constrained proliferation, migration, invasion, and EMT(epithel-ial-mesenchymal transition), and facilitated apoptosis of HCC cells in vitro.Knockdo-wn of circBACH1 reduced HCC growth in vivo. SERBP1 overexpression partially neutralized the repressive effect of circBACH1 silencing on malignant behaviors of HCC cells.CircBACH1 sponged miR-656-3p to elevate SERBP1 expression, thereby accelerating the progression of HCC.The research provided a new evidence to support the role of circBACH1 in HCC.
Assuntos
Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Progressão da Doença , Neoplasias Hepáticas , MicroRNAs/genética , RNA Circular/genética , Proteínas de Ligação a RNA/genética , Animais , Linhagem Celular Tumoral , Proliferação de Células , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Técnicas In Vitro , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Camundongos , Camundongos Endogâmicos BALB C , RNA Circular/deficiência , Proteínas de Ligação a RNA/antagonistas & inibidores , Proteínas de Ligação a RNA/biossíntese , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Circular RNAs (circRNAs) and microRNAs (miRNAs) have been emerging as new players in acute myeloid leukemia (AML). Hsa_circ_0005774 (circ_0005774) is an upregulated circRNA in pediatric AML, while its role is uncovered. Thus, we intended to measure the function and mechanism of circ_0005774 in AML leukemogenesis. Real time-quantitative PCR revealed that circ_0005774 was highly expressed in blood of pediatric AML patients and AML cells (HL-60 and NB4), accompanied with downregulated miRNA-192-5p (miR-192-5p) which was a crucial tumor-associated and leukemia-related miRNA. Circ_0005774 was abundant in miRNA response element according to CSCD software, and miR-192-5p was identified as a target of circ_0005774, as evidenced by RNA immunoprecipitation and dual-luciferase reporter assays. Cell viability assay, flow cytometry and western blotting were performed to measure cell functions. Accordingly, blocking circ_0005774 and/or overexpressing miR-192-5p could enhance apoptosis rate of HL-60 and NB4 cells, but suppress cell viability and cell cycle entrance, accompanied with depression of proliferation markers including proliferating cell nuclear antigen (PCNA), CyclinD1 and B cell lymphoma 2 (Bcl-2). Meanwhile, depleting miR-192-5p counteracted the role of circ_0005774 knockdown in AML cells. Uncoordinated 51-like kinase 1 (ULK1) was previously demonstrated to be associated with diagnosis, prognosis and therapeutic strategy for AML, and restoring ULK1 could abrogate miR-192-5p overexpression-induced effects in HL-60 and NB4 cells. Notably, ULK1 was a downstream target of miR-192-5p and indirectly modulated by circ_0005774. In conclusion, circ_0005774 knockdown repressed cell proliferation and promoted apoptosis of AML cells partially through regulating miR-192-5p/ULK1 axis.
Assuntos
Apoptose , Proteína Homóloga à Proteína-1 Relacionada à Autofagia/genética , Peptídeos e Proteínas de Sinalização Intracelular/genética , Leucemia Mieloide Aguda/sangue , Leucemia Mieloide Aguda/patologia , MicroRNAs/genética , RNA Circular/sangue , RNA Circular/genética , Proteína Homóloga à Proteína-1 Relacionada à Autofagia/metabolismo , Sequência de Bases , Linhagem Celular Tumoral , Proliferação de Células , Sobrevivência Celular , Criança , Pré-Escolar , Feminino , Técnicas de Silenciamento de Genes , Inativação Gênica , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/metabolismo , Masculino , RNA Circular/deficiência , Regulação para CimaRESUMO
BACKGROUND: Circular RNAs (circRNAs) play an important role in the tumorigenesis of pancreatic cancer. However, the expression profiles and roles of circRNAs in pancreatic cancer remain largely unknown. METHODS: To identify differentially expressed circRNAs (DEcircRNAs) between pancreatic cancer and matched normal tissues, bioinformatics analysis was performed. Hsa_circ_0000069 was identified by 0.bioinformatics analysis. In addition, the level of hsa_circ_0000069 in pancreatic cancer tissues and cell lines, and pancreatic cancer cell-derived exosomes were assessed using RT-qPCR assay. RESULTS: The expression of hsa_circ_0000069 was markedly upregulated in pancreatic cancer tissues and cell lines. SCL/TAL1 interrupting locus (STIL) is the parent gene for hsa_circ_0000069, and its high expression was related to poor overall survival in patients with pancreatic cancer. In addition, downregulation of hsa_circ_0000069 markedly suppressed STIL expression, induced the apoptosis and cell cycle arrest, and inhibited the proliferation, migration and invasion in pancreatic cancer cells. Moreover, hsa_circ_0000069 knockdown inhibited the growth of xenograft pancreatic cancer tumors in vivo. Furthermore, human pancreatic duct epithelial cells (HPDE) are capable of internalizing SW1990 cell-derived exosomes, allowing the transfer of hsa_circ_0000069. Significantly, SW1990 cell-derived exosomes promoted the proliferation, migration and cell cycle progression of HPDE cells, whereas exosomes with downregulated hsa_circ_0000069 suppressed the proliferation, migration and cell cycle progression of HPDE cells, by suppressing STIL expression. CONCLUSION: Our results suggest that hsa_circ_0000069 knockdown could inhibit pancreatic cancer tumorigenesis and exosomes with downregulated hsa_circ_0000069 could suppress HPDE cell malignant transformation. Collectively, hsa_circ_0000069 might be a therapeutic target for the treatment of pancreatic cancer.
Assuntos
Carcinogênese/genética , Transformação Celular Neoplásica/genética , Exossomos/genética , Técnicas de Silenciamento de Genes , Peptídeos e Proteínas de Sinalização Intracelular/genética , Neoplasias Pancreáticas/patologia , RNA Circular/genética , Apoptose/genética , Pontos de Checagem do Ciclo Celular/genética , Linhagem Celular Tumoral , Proliferação de Células/genética , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Neoplasias Pancreáticas/genética , RNA Circular/deficiênciaRESUMO
Diabetic kidney disease (DKD) is the leading cause of chronic kidney disease. Current therapies for DKD are insufficient. Therefore, there is an urgent need for identifying new therapies. An increasing number of micro RNAs (miRNAs) and long noncoding RNAs (lncRNAs) have been demonstrated to modulate the progression of diabetic kidney disease. Nevertheless, until now, there have been few reports evaluating the relevance of circular RNAs (circRNAs) in DKD. circRNAs have been reported to regulate the occurrence and development of multiple diseases. In this study, we intended to explore the circRNA expression profiles and determine the role of circRNA in DKD. We identified a series of dysregulated circRNAs in glucose-stressed HK-2 cells using circRNA microarray analysis. Among the candidate circRNAs, we found that circACTR2 was upregulated and may be involved in inflammation and pyroptosis. Knockdown of circACTR2 significantly decreased pyroptosis, interleukin (IL)-1ß release and collagen IV and fibronectin production, indicating the effective regulation by circACTR2 of cell death and inflammation. Overall, our study identified a new circRNA, circACTR2, that regulates high glucose-induced pyroptosis, inflammation and fibrosis in proximal tubular cells. The present study preliminarily explores the role of circRNAs in pyroptosis of tubular cells, and provides novel insight into the pathogenesis of DKD and new therapeutic strategies.
Assuntos
Nefropatias Diabéticas/genética , Nefropatias Diabéticas/metabolismo , Glucose/efeitos adversos , Piroptose/genética , Piroptose/fisiologia , RNA Circular/deficiência , RNA Circular/genética , Proteína 2 Relacionada a Actina , Linhagem Celular , Colágeno/metabolismo , Células Epiteliais , Fibronectinas/metabolismo , Fibrose/genética , Fibrose/metabolismo , Expressão Gênica , Humanos , Interleucina-1beta/metabolismo , Rim/metabolismoRESUMO
Background: Prostate cancer (PC) fearfully impacts men's health. We explored the efficacy and mechanism of circular RNA circZNF609 (circZNF609) on colony formation, viability, apoptosis, migration and invasion and in PC cells. Methods: Colony formation, CCK-8, flow cytometry, migration and invasion assay were respectively used to detect the functions of circZNF609 and microRNA (miR)-186-5p on cell colony ability, viability, apoptosis, migration and invasion. circZNF609 and miR-186-5p expression were changed by cell transfection and tested by RT-qPCR. Moreover, Cleaved-Caspase-3, Cleaved-Caspase-9, matrix metalloproteinase-9 (MMP-9), Vimentin and relate-proteins of cell pathways were examined through Western blot. Results: circZNF609 was highly expressed at PC tissues. circZNF609 declined cell colony ability, viability, migration and invasion and caused apoptosis. Furthermore, circZNF609 negatively regulated miR-186-5p, miR-186-5p inhibitor could reverse impacts of circZNF609. Finally, circZNF609 restrained the YAP1 and AMPK pathways by up-regulating miR-186-5p. Conclusion: Silencing circZNF609 restrained growth, migration and invasion of PC cells by up-regulating miR-186-5p via YAP1 and AMPK pathways. Highlights circZNF609 is highly expressed in PC tissues; circZNF609 restrains cell growth, migration and invasion; circZNF609 exerts its function by up-regulating miR-186-5p; circZNF609 exerts its function by YAP1 and AMPK signaling pathways.