ABSTRACT
BACKGROUND: Mounting evidence has shown that circular RNAs (circRNAs) have vital roles in human cancers, including retinoblastoma (RB). The purpose of this study was to investigate the exact roles and underlying mechanism of circRNA ER membrane protein complex subunit 9 (circ-FAM158A) in RB. METHODS: Quantitative real-time polymerase chain reaction (qRT-PCR) was carried out to determine the expression levels of circ-FAM158A, miR-138-5p and solute carrier family 7 member 5 (SLC7A5). Cell proliferation was evaluated by Cell counting Kit-8 (CCK-8) assay and colony formation assay. Flow cytometry analysis was applied to determine cell cycle distribution and apoptosis rate. Transwell assay was conducted to assess cell migration and invasion. The interaction between miR-138-5p and circ-FAM158A or SLC7A5 was predicted by starBase v2.0 and confirmed by dual-luciferase reporter assay. Western blot assay was performed to examine the protein expression of SLC7A5. The mice xenograft model was established, immunohistochemistry (IHC) and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labelling (TUNEL) assays were conducted to confirm the role of circ-FAM158A in vivo. RESULTS: Circ-FAM158A and SLC7A5 were overexpressed and miR-138-5p was downregulated in RB tissues and cells. Circ-FAM158A knockdown inhibited RB cell proliferation, metastasis, and promoted apoptosis in vitro and in vivo. MiR-138-5p was a direct target of circ-FAM158A, and miR-138-5p inhibition reversed the inhibitory effect of circ-FAM158A silence on the progression of RB cells. Additionally, SLC7A5 was identified as a target of miR-138-5p, and SLC7A5 overexpression abated the anti-tumor roles of miR-138-5p in RB cells. Besides, circ-FAM158A positively regulated SLC7A5 expression by sponging miR-138-5p. CONCLUSION: Circ-FAM158A knockdown inhibited the progression of RB by regulating miR-138-5p/SLC7A5 axis, which provided new insights into the pathogenesis of RB.
Subject(s)
Gene Expression Regulation, Neoplastic/physiology , Large Neutral Amino Acid-Transporter 1/genetics , Membrane Proteins/genetics , MicroRNAs/genetics , RNA, Circular/genetics , Retinal Neoplasms/genetics , Retinoblastoma/genetics , Animals , Apoptosis , Blotting, Western , Cell Movement , Cell Proliferation , Disease Progression , Female , Flow Cytometry , Gene Silencing , Heterografts , Humans , Immunohistochemistry , In Situ Nick-End Labeling , Mice , Mice, Inbred BALB C , Mice, Nude , Real-Time Polymerase Chain Reaction , Retinal Neoplasms/pathology , Retinoblastoma/pathology , Transfection , Tumor Cells, Cultured , Up-RegulationABSTRACT
AIMS OR PURPOSE: Glaucoma is the leading cause of vision loss and blindness in the world. Elucidating the pathogenesis of glaucoma and developing effective treatment should be the priority. Inflammation and oxidative stress play essential roles in glaucoma pathogeneisis. Kaempferol is a natural flavonol and has anti-inflammatory and anti-oxidative activities. In this study, we explored the potential effects of kaempferol on acute glaucoma. METHODS: We established the retinal ischemia-reperfusion (I/R) mice model and administrated kaempferol to I/R mice. We monitored the retina thickness change, retinal ganglion cell (RGC) death, caspase-8 and caspase-3 activation, NLRP1/NLRP3 inflammasomes activation, pro-inflammatory cytokines production, and activations of NF-κB and MAPKs signaling pathways. RESULTS: Kaempferol prevented retina thickness change and RGC death in I/R mice. The activations of caspase-8, caspase-3, and NLRP1/NLRP3 inflammasome activation were inhibited by kaempferol. Kaempferol prevented pro-inflammatory cytokines productions in I/R mice. The activation of NF-κB and JNK signaling pathways was also inhibited by Kaempferol in I/R mice. CONCLUSION: Kaempferol attenuated retinal ganglion cell death by suppressing NLRP1/NLRP3 inflammasomes and caspase-8 via inhibiting NF-κB and JNK pathways in acute glaucoma.
Subject(s)
Adaptor Proteins, Signal Transducing/metabolism , Apoptosis Regulatory Proteins/metabolism , Caspase 8/metabolism , Inflammasomes/metabolism , JNK Mitogen-Activated Protein Kinases/metabolism , Kaempferols/pharmacology , NF-kappa B p50 Subunit/metabolism , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , Retinal Ganglion Cells/drug effects , Animals , Antioxidants/pharmacology , Blotting, Western , Cell Survival/drug effects , Disease Models, Animal , Glaucoma/drug therapy , Glaucoma/metabolism , Intraocular Pressure/drug effects , Mice, Inbred C57BL , Microscopy, Fluorescence , Real-Time Polymerase Chain Reaction , Reperfusion Injury , Retinal Ganglion Cells/metabolismABSTRACT
PURPOSE: To investigate the biochemical characteristics in experimental keratomycosis by Raman spectroscopy analysis in vitro and in vivo. METHODS: Raman spectroscopy was used to analyze the biochemical characteristics of cultured mouse keratocytes stimulated by Fusarium solani suspension in vitro, and the infected cornea of Fusarium solani keratitis of mice. RESULTS: The peak intensities at 1005, 1186, 1311, 1449 and 1657 cm-1, which represented phenylalanine, tyrosine, nucleic acid bases, phospholipids and α-helix, were decreased in the infected keratocytes compared with the control keratocytes. The consistency of Raman spectra between the infected cornea tissue and the control cornea tissue was high. However, the intensity of some peaks declined, especially at 853, 940 and 1244 cm-1, which represent the tyrosine, proline and collagen content, respectively. CONCLUSIONS: After infection with Fusarium solani, the biochemical characteristics of keratocyte and cornea showed a decrease of amino acids, nucleic acid phospholipids and collagen, which may closely relate to the pathophysiology of keratomycosis.
