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1.
Nucleic Acids Res ; 50(21): 12235-12250, 2022 11 28.
Article in English | MEDLINE | ID: mdl-36477888

ABSTRACT

Neural differentiation of embryonic stem cells (ESCs) requires precisely orchestrated gene regulation, a process governed in part by changes in 3D chromatin structure. How these changes regulate gene expression in this context remains unclear. In this study, we observed enrichment of the transcription factor KLF4 at some poised or closed enhancers at TSS-linked regions of genes associated with neural differentiation. Combination analysis of ChIP, HiChIP and RNA-seq data indicated that KLF4 loss in ESCs induced changes in 3D chromatin structure, including increased chromatin interaction loops between neural differentiation-associated genes and active enhancers, leading to upregulated expression of neural differentiation-associated genes and therefore early neural differentiation. This study suggests KLF4 inhibits early neural differentiation by regulation of 3D chromatin structure, which is a new mechanism of early neural differentiation.


Subject(s)
Chromatin , Embryonic Stem Cells , Kruppel-Like Factor 4 , Cell Differentiation/genetics , Chromatin/metabolism , Embryonic Stem Cells/metabolism , Gene Expression Regulation , Transcription Factors/metabolism , Kruppel-Like Factor 4/metabolism
2.
Genome Res ; 30(2): 155-163, 2020 02.
Article in English | MEDLINE | ID: mdl-31953347

ABSTRACT

Temozolomide (TMZ) is a frequently used chemotherapy for glioma; however, chemoresistance is a major problem limiting its effectiveness. Thus, knowledge of mechanisms underlying this outcome could improve patient prognosis. Here, we report that deletion of a regulatory element in the HOTAIR locus increases glioma cell sensitivity to TMZ and alters transcription of multiple genes. Analysis of a combination of RNA-seq, Capture Hi-C, and patient survival data suggests that CALCOCO1 and ZC3H10 are target genes repressed by the HOTAIR regulatory element and that both function in regulating glioma cell sensitivity to TMZ. Rescue experiments and 3C data confirmed this hypothesis. We propose a new regulatory mechanism governing glioma cell TMZ sensitivity.


Subject(s)
Calcium-Binding Proteins/genetics , Carrier Proteins/genetics , Glioma/drug therapy , RNA, Long Noncoding/genetics , Temozolomide/pharmacology , Transcription Factors/genetics , Antineoplastic Agents, Alkylating/pharmacology , Base Sequence , CRISPR-Cas Systems/genetics , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Survival/drug effects , Drug Resistance, Neoplasm/genetics , Gene Expression Regulation, Neoplastic/drug effects , Gene Regulatory Networks/genetics , Glioma/genetics , Glioma/pathology , Humans , Neoplasm Proteins/genetics
3.
Nucleic Acids Res ; 49(20): 11575-11595, 2021 11 18.
Article in English | MEDLINE | ID: mdl-34723340

ABSTRACT

Signaling pathway-driven target gene transcription is critical for fate determination of embryonic stem cells (ESCs), but enhancer-dependent transcriptional regulation in these processes remains poorly understood. Here, we report enhancer architecture-dependent multilayered transcriptional regulation at the Halr1-Hoxa1 locus that orchestrates retinoic acid (RA) signaling-induced early lineage differentiation of ESCs. We show that both homeobox A1 (Hoxa1) and Hoxa adjacent long non-coding RNA 1 (Halr1) are identified as direct downstream targets of RA signaling and regulated by RARA/RXRA via RA response elements (RAREs). Chromosome conformation capture-based screens indicate that RA signaling promotes enhancer interactions essential for Hoxa1 and Halr1 expression and mesendoderm differentiation of ESCs. Furthermore, the results also show that HOXA1 promotes expression of Halr1 through binding to enhancer; conversely, loss of Halr1 enhances interaction between Hoxa1 chromatin and four distal enhancers but weakens interaction with chromatin inside the HoxA cluster, leading to RA signaling-induced Hoxa1 overactivation and enhanced endoderm differentiation. These findings reveal complex transcriptional regulation involving synergistic regulation by enhancers, transcription factors and lncRNA. This work provides new insight into intrinsic molecular mechanisms underlying ESC fate determination during RA signaling-induced early differentiation.


Subject(s)
Cell Differentiation , Enhancer Elements, Genetic , Mouse Embryonic Stem Cells/metabolism , Tretinoin/pharmacology , Animals , Cell Line , Cell Lineage , Cells, Cultured , Chromatin Assembly and Disassembly , Homeodomain Proteins/genetics , Homeodomain Proteins/metabolism , Mice , Mouse Embryonic Stem Cells/cytology , Mouse Embryonic Stem Cells/drug effects , RNA, Long Noncoding/genetics , RNA, Long Noncoding/metabolism , Signal Transduction , Transcription Factors/genetics , Transcription Factors/metabolism
4.
Int J Mol Sci ; 24(19)2023 Oct 06.
Article in English | MEDLINE | ID: mdl-37834393

ABSTRACT

Ferroptosis is an iron-dependent form of cell death, which is reported to be associated with glioma progression and drug sensitivity. Targeting ferroptosis is a potential therapeutic approach for glioma. However, the molecular mechanism of glioma cell ferroptosis is not clear. In this study, we profile the change of 3D chromatin structure in glioblastoma ferroptosis by using HiChIP and study the 3D gene regulation network in glioblastoma ferroptosis. A combination of an analysis of HiChIP and RNA-seq data suggests that change of chromatin loops mediated by 3D chromatin structure regulates gene expressions in glioblastoma ferroptosis. Genes that are regulated by 3D chromatin structures include genes that were reported to function in ferroptosis, like HDM2 and TXNRD1. We propose a new regulatory mechanism governing glioblastoma cell ferroptosis by 3D chromatin structure.


Subject(s)
Ferroptosis , Glioblastoma , Glioma , Humans , Glioblastoma/genetics , Ferroptosis/genetics , Cell Death , Chromatin/genetics
5.
J Biol Chem ; 296: 100413, 2021.
Article in English | MEDLINE | ID: mdl-33581110

ABSTRACT

Proper expression of Homeobox A cluster genes (HoxA) is essential for embryonic stem cell (ESC) differentiation and individual development. However, mechanisms controlling precise spatiotemporal expression of HoxA during early ESC differentiation remain poorly understood. Herein, we identified a functional CTCF-binding element (CBE+47) closest to the 3'-end of HoxA within the same topologically associated domain (TAD) in ESC. CRISPR-Cas9-mediated deletion of CBE+47 significantly upregulated HoxA expression and enhanced early ESC differentiation induced by retinoic acid (RA) relative to wild-type cells. Mechanistic analysis by chromosome conformation capture assay (Capture-C) revealed that CBE+47 deletion decreased interactions between adjacent enhancers, enabling formation of a relatively loose enhancer-enhancer interaction complex (EEIC), which overall increased interactions between that EEIC and central regions of HoxA chromatin. These findings indicate that CBE+47 organizes chromatin interactions between its adjacent enhancers and HoxA. Furthermore, deletion of those adjacent enhancers synergistically inhibited HoxA activation, suggesting that these enhancers serve as an EEIC required for RA-induced HoxA activation. Collectively, these results provide new insight into RA-induced HoxA expression during early ESC differentiation, also highlight precise regulatory roles of the CTCF-binding element in orchestrating high-order chromatin structure.


Subject(s)
CCCTC-Binding Factor/metabolism , Embryonic Stem Cells/metabolism , Homeodomain Proteins/metabolism , Animals , CCCTC-Binding Factor/physiology , Cell Differentiation , Cell Line , Chromatin/genetics , Chromatin/metabolism , Chromatin Assembly and Disassembly , Embryonic Stem Cells/physiology , Enhancer Elements, Genetic/genetics , Gene Expression/genetics , Gene Expression Regulation/genetics , Genes, Homeobox/genetics , Homeodomain Proteins/genetics , Homeodomain Proteins/physiology , Mice , Transcriptional Activation , Tretinoin/pharmacology
6.
Nucleic Acids Res ; 47(13): 6737-6752, 2019 07 26.
Article in English | MEDLINE | ID: mdl-31147716

ABSTRACT

Retinoic acid (RA) induces rapid differentiation of embryonic stem cells (ESCs), partly by activating expression of the transcription factor Hoxa1, which regulates downstream target genes that promote ESCs differentiation. However, mechanisms of RA-induced Hoxa1 expression and ESCs early differentiation remain largely unknown. Here, we identify a distal enhancer interacting with the Hoxa1 locus through a long-range chromatin loop. Enhancer deletion significantly inhibited expression of RA-induced Hoxa1 and endoderm master control genes such as Gata4 and Gata6. Transcriptome analysis revealed that RA-induced early ESCs differentiation was blocked in Hoxa1 enhancer knockout cells, suggesting a requirement for the enhancer. Restoration of Hoxa1 expression partly rescued expression levels of ∼40% of genes whose expression changed following enhancer deletion, and ∼18% of promoters of those rescued genes were directly bound by Hoxa1. Our data show that a distal enhancer maintains Hoxa1 expression through long-range chromatin loop and that Hoxa1 directly regulates downstream target genes expression and then orchestrates RA-induced early differentiation of ESCs. This discovery reveals mechanisms of a novel enhancer regulating RA-induced Hoxa genes expression and early ESCs differentiation.


Subject(s)
Embryonic Stem Cells/metabolism , Enhancer Elements, Genetic , Homeodomain Proteins/biosynthesis , Transcription Factors/biosynthesis , Tretinoin/pharmacology , Animals , CRISPR-Cas Systems , Cell Differentiation/drug effects , Cells, Cultured , Chromatin/genetics , Chromatin/metabolism , Embryonic Stem Cells/drug effects , Endoderm/metabolism , Enhancer Elements, Genetic/genetics , Gene Editing , Gene Expression Regulation, Developmental , Gene Knockdown Techniques , Gene Ontology , Homeodomain Proteins/genetics , Intracellular Signaling Peptides and Proteins/biosynthesis , Intracellular Signaling Peptides and Proteins/genetics , Mice , Promoter Regions, Genetic , RNA, Small Interfering/genetics , Recombinant Proteins/metabolism , Transcription Factors/genetics
7.
Int J Mol Sci ; 22(15)2021 Aug 03.
Article in English | MEDLINE | ID: mdl-34361106

ABSTRACT

Enhancers regulate multiple genes via higher-order chromatin structures, and they further affect cancer progression. Epigenetic changes in cancer cells activate several cancer-specific enhancers that are silenced in normal cells. These cancer-specific enhancers are potential therapeutic targets of cancer. However, the functions and regulation networks of colorectal-cancer-specific enhancers are still unknown. In this study, we profile colorectal-cancer-specific enhancers and reveal their regulation network through the analysis of HiChIP data that were derived from a colorectal cancer cell line and Hi-C and RNA-seq data that were derived from tissue samples by in silico analysis and in vitro experiments. Enhancer-promoter loops in colorectal cancer cells containing colorectal-cancer-specific enhancers are involved in more than 50% of the topological associated domains (TADs) changed in colorectal cancer cells compared to normal colon cells. In addition, colorectal-cancer-specific enhancers interact with 152 genes that are significantly and highly expressed in colorectal cancer cells. These colorectal-cancer-specific enhancer target genes include ITGB4, RECQL4, MSLN, and GDF15. We propose that the regulation network of colorectal-cancer-specific enhancers plays an important role in the progression of colorectal cancer.


Subject(s)
Biomarkers, Tumor/metabolism , Colorectal Neoplasms/pathology , Enhancer Elements, Genetic , Epigenesis, Genetic , Gene Expression Regulation, Neoplastic , Promoter Regions, Genetic , Apoptosis , Biomarkers, Tumor/genetics , Cell Proliferation , Colorectal Neoplasms/genetics , Disease Progression , Humans , Mesothelin , Tumor Cells, Cultured
8.
Mol Biol Rep ; 47(4): 2723-2733, 2020 Apr.
Article in English | MEDLINE | ID: mdl-32180085

ABSTRACT

The long noncoding RNA HOTAIRM1 reportedly plays important roles in acute myeloid leukemia, gastric cancer and colorectal cancer. Here, we analyzed potential function of HOTAIRM1 in glioma and asked whether it participates in long-range chromatin interactions. We monitored expression of HOTAIRM1 in glioma tissues and correlated levels with patient survival using the TCGA dataset. HOTAIRM1 was highly expressed in glioma tissue, with high levels associated with shortened patient survival time. We then suppressed HOTAIRM1 activity in the human glioblastoma U251 line using CRISPR-cas9 to knock in a truncating polyA fragment. Reporter analysis of these and control cells confirmed that the HOTAIRM1 locus serves as an active enhancer. We then performed Capture-C analysis to identify target genes of that locus and applied RNA antisense purification to assess chromatin interactions between the HOTAIRM1 locus and HOXA cluster genes. HOTAIRM1 knockdown in glioma cells decreased proliferation and reduced expression of HOXA cluster genes. HOTAIRM1 regulates long-range interactions between the HOTAIRM1 locus and HOXA genes. Our work suggests a new mechanism by which HOTAIRM1 regulates glioma progression by regulating high-order chromatin structure and could suggest novel therapeutic targets to treat an intractable cancer.


Subject(s)
Brain Neoplasms/genetics , Glioma/genetics , Homeodomain Proteins/genetics , MicroRNAs/genetics , Brain Neoplasms/metabolism , Brain Neoplasms/pathology , Cell Line, Tumor , Cell Proliferation/physiology , Chromatin/genetics , Chromatin/metabolism , Databases, Genetic , Gene Expression Profiling , Glioma/metabolism , Glioma/pathology , Homeodomain Proteins/metabolism , Humans , MicroRNAs/metabolism , Multigene Family , RNA, Long Noncoding/genetics
9.
Proc Natl Acad Sci U S A ; 114(50): E10717-E10725, 2017 12 12.
Article in English | MEDLINE | ID: mdl-29180410

ABSTRACT

The receptor-like tyrosine kinase (Ryk), a Wnt receptor, is important for cell fate determination during corticogenesis. During neuronal differentiation, the Ryk intracellular domain (ICD) is cleaved. Cleavage of Ryk and nuclear translocation of Ryk-ICD are required for neuronal differentiation. However, the mechanism of translocation and how it regulates neuronal differentiation remain unclear. Here, we identified Smek1 and Smek2 as Ryk-ICD partners that regulate its nuclear localization and function together with Ryk-ICD in the nucleus through chromatin recruitment and gene transcription regulation. Smek1/2 double knockout mice displayed pronounced defects in the production of cortical neurons, especially interneurons, while the neural stem cell population increased. In addition, both Smek and Ryk-ICD bound to the Dlx1/2 intergenic regulator element and were involved in its transcriptional regulation. These findings demonstrate a mechanism of the Ryk signaling pathway in which Smek1/2 and Ryk-ICD work together to mediate neural cell fate during corticogenesis.


Subject(s)
Molecular Chaperones/metabolism , Neurogenesis/physiology , Phosphoprotein Phosphatases/metabolism , Receptor Protein-Tyrosine Kinases/metabolism , Animals , Cell Nucleus/metabolism , Cells, Cultured , Coenzymes/metabolism , HEK293 Cells , Humans , Mice
11.
J Biol Chem ; 288(42): 30399-30410, 2013 Oct 18.
Article in English | MEDLINE | ID: mdl-24005670

ABSTRACT

Abnormal osteoclast formation and osteolysis are the hallmarks of multiple myeloma (MM) bone disease, yet the underlying molecular mechanisms are incompletely understood. Here, we show that the AKT pathway was up-regulated in primary bone marrow monocytes (BMM) from patients with MM, which resulted in sustained high expression of the receptor activator of NF-κB (RANK) in osteoclast precursors. The up-regulation of RANK expression and osteoclast formation in the MM BMM cultures was blocked by AKT inhibition. Conditioned media from MM cell cultures activated AKT and increased RANK expression and osteoclast formation in BMM cultures. Inhibiting AKT in cultured MM cells decreased their growth and ability to promote osteoclast formation. Of clinical significance, systemic administration of the AKT inhibitor LY294002 blocked the formation of tumor tissues in the bone marrow cavity and essentially abolished the MM-induced osteoclast formation and osteolysis in SCID mice. The level of activating transcription factor 4 (ATF4) protein was up-regulated in the BMM cultures from multiple myeloma patients. Adenoviral overexpression of ATF4 activated RANK expression in osteoclast precursors. These results demonstrate a new role of AKT in the MM promotion of osteoclast formation and bone osteolysis through, at least in part, the ATF4-dependent up-regulation of RANK expression in osteoclast precursors.


Subject(s)
Gene Expression Regulation, Neoplastic , Multiple Myeloma/enzymology , Osteoclasts/enzymology , Osteolysis/enzymology , Proto-Oncogene Proteins c-akt/metabolism , Up-Regulation , Activating Transcription Factor 4/metabolism , Animals , Chromones/pharmacology , Enzyme Inhibitors/pharmacology , Female , Heterografts , Humans , Male , Mice , Mice, SCID , Morpholines/pharmacology , Multiple Myeloma/pathology , Neoplasm Transplantation , Osteoclasts/pathology , Osteolysis/pathology , Proto-Oncogene Proteins c-akt/antagonists & inhibitors , Receptor Activator of Nuclear Factor-kappa B/metabolism , Tumor Cells, Cultured
12.
Cell Death Dis ; 15(1): 6, 2024 01 04.
Article in English | MEDLINE | ID: mdl-38177123

ABSTRACT

Glioma cell sensitivity to temozolomide (TMZ) is critical for effective treatment and correlates with patient survival, although mechanisms underlying this activity are unclear. Here, we reveal a new mechanism used by glioma cells to modulate TMZ sensitivity via regulation of SORBS2 and DDR1 genes by super-enhancer RNA LINC02454. We report that LINC02454 activity increases glioma cell TMZ sensitivity by maintaining long-range chromatin interactions between SORBS2 and the LINC02454 enhancer. By contrast, LINC02454 activity also decreased glioma cell TMZ sensitivity by promoting DDR1 expression. Our study suggests a bivalent function for super-enhancer RNA LINC02454 in regulating glioma cell sensitivity to TMZ.


Subject(s)
Brain Neoplasms , Glioma , MicroRNAs , Humans , Temozolomide/pharmacology , Temozolomide/therapeutic use , Enhancer RNAs , Drug Resistance, Neoplasm/genetics , Cell Line, Tumor , Glioma/drug therapy , Glioma/genetics , Glioma/metabolism , MicroRNAs/genetics , Cell Proliferation , Brain Neoplasms/drug therapy , Brain Neoplasms/genetics , Brain Neoplasms/metabolism , Antineoplastic Agents, Alkylating/pharmacology , Antineoplastic Agents, Alkylating/therapeutic use
13.
iScience ; 26(4): 106497, 2023 Apr 21.
Article in English | MEDLINE | ID: mdl-37096036

ABSTRACT

To date, genome-wide association studies (GWAS) have revealed over 200 genetic risk loci associated with prostate cancer; yet, true disease-causing variants remain elusive. Identification of causal variants and their targets from association signals is complicated by high linkage disequilibrium and limited availability of functional genomics data for specific tissue/cell types. Here, we integrated statistical fine-mapping and functional annotation from prostate-specific epigenomic profiles, 3D genome features, and quantitative trait loci data to distinguish causal variants from associations and identify target genes. Our fine-mapping analysis yielded 3,395 likely causal variants, and multiscale functional annotation linked them to 487 target genes. We prioritized rs10486567 as a genome-wide top-ranked SNP and predicted HOTTIP as its target. Deletion of the rs10486567-associated enhancer in prostate cancer cells decreased their capacity for invasive migration. HOTTIP overexpression in enhancer-KO cell lines rescued defective invasive migration. Furthermore, we found that rs10486567 regulates HOTTIP through allele-specific long-range chromatin interaction.

14.
J Am Acad Dermatol ; 67(6): 1210-3, 2012 Dec.
Article in English | MEDLINE | ID: mdl-22516113

ABSTRACT

BACKGROUND: "Wait-and-see" is a common principle for most superficial infantile hemangiomas (IHs) because of their expected involution. Topical propranolol has recently been reported to be an effective treatment for superficial IHs. OBJECTIVE: The aim of this study was to evaluate the efficacy and safety of 1% propranolol ointment in the treatment of superficial IHs. METHODS: A retrospective chart review was performed on 25 children (21 female and 4 male) with a median age of 4 months (range, 1-10 months). A total of 28 lesions were treated with 1% propranolol ointment. Topical propranolol was applied thrice daily for a mean duration of 21 weeks (range, 5-59 weeks). Changes in the size, texture, and color of the tumor were monitored and recorded at regular intervals. The treatment response was evaluated using a 3-point scale system: good, partial, and no response. Adverse effects after medication were evaluated and managed accordingly. RESULTS: Of the 28 hemangiomas, 16 (57%) demonstrated good response, 9 (33%) showed a partial response, and 3 (10%) had no response. Among all the IHs, 90% showed either good or partial responses to topical 1% propranolol ointment treatment. No systemic complication was observed in any of the patients. LIMITATIONS: This report is a retrospective uncontrolled study. CONCLUSIONS: Topical therapy with 1% propranolol ointment may be a safe and effective method for the treatment of superficial IHs and can be used as an adjuvant treatment measure during the wait-and-see period.


Subject(s)
Adrenergic beta-Antagonists/administration & dosage , Hemangioma/drug therapy , Propranolol/administration & dosage , Skin Neoplasms/drug therapy , Administration, Topical , Female , Humans , Infant , Male , Ointments , Retrospective Studies
15.
Sci Rep ; 12(1): 18678, 2022 11 04.
Article in English | MEDLINE | ID: mdl-36333421

ABSTRACT

Lichen planus (LP) is a chronic inflammatory disease. Oral lichen planus (OLP) mainly appears as oral mucosal reticular or ulcerative lesions with an unknown etiology. We aimed to explore the immunomodulatory effect of paeoniflorin (PF) in mesenchymal stem cells (MSCs) and the potential involvement of Th1/Th2 cytokines in OLP. The effects of paeoniflorin on the proliferation and migration of MSCs were detected by Cell Counting Kit-8 (CCK8) and Transwell assays. MSCs were subjected to osteogenic, adipogenic and neurogenic induction followed by Alizarin red, oil red O, real-time PCR and immunofluorescence assays. We found that paeoniflorin promoted the proliferation, migration and multilineage differentiation of MSCs from OLP lesions (OLP-MSCs) in vitro. Paeoniflorin pretreatment increased the inhibitory effect of OLP-MSCs on peripheral blood mononuclear cells. Furthermore, paeoniflorin-pretreated OLP-MSCs simultaneously decreased Th1 cytokine levels and increased Th2 cytokine levels in T lymphocyte cocultures. Finally, paeoniflorin-pretreated OLP-MSCs also promoted the Th1/Th2 balance both in vitro and in the serum of mice that received skin allografts. In conclusion, paeoniflorin enhanced MSC immunomodulation and changed the inflammatory microenvironment via T lymphocytes, suggesting that the improvement of OLP-MSCs is a promising therapeutic approach for OLP.


Subject(s)
Lichen Planus, Oral , Mesenchymal Stem Cells , Mice , Animals , Lichen Planus, Oral/pathology , Cytokines , Leukocytes, Mononuclear/pathology , Mesenchymal Stem Cells/pathology , Immunomodulation
16.
Eye Vis (Lond) ; 9(1): 13, 2022 Apr 01.
Article in English | MEDLINE | ID: mdl-35361278

ABSTRACT

BACKGROUND: Myopic maculopathy (MM) has become a major cause of visual impairment and blindness worldwide, especially in East Asian countries. Deep learning approaches such as deep convolutional neural networks (DCNN) have been successfully applied to identify some common retinal diseases and show great potential for the intelligent analysis of MM. This study aimed to build a reliable approach for automated detection of MM from retinal fundus images using DCNN models. METHODS: A dual-stream DCNN (DCNN-DS) model that perceives features from both original images and corresponding processed images by color histogram distribution optimization method was designed for classification of no MM, tessellated fundus (TF), and pathologic myopia (PM). A total of 36,515 gradable images from four hospitals were used for DCNN model development, and 14,986 gradable images from the other two hospitals for external testing. We also compared the performance of the DCNN-DS model and four ophthalmologists on 3000 randomly sampled fundus images. RESULTS: The DCNN-DS model achieved sensitivities of 93.3% and 91.0%, specificities of 99.6% and 98.7%, areas under the receiver operating characteristic curves (AUC) of 0.998 and 0.994 for detecting PM, whereas sensitivities of 98.8% and 92.8%, specificities of 95.6% and 94.1%, AUCs of 0.986 and 0.970 for detecting TF in two external testing datasets. In the sampled testing dataset, the sensitivities of four ophthalmologists ranged from 88.3% to 95.8% and 81.1% to 89.1%, and the specificities ranged from 95.9% to 99.2% and 77.8% to 97.3% for detecting PM and TF, respectively. Meanwhile, the DCNN-DS model achieved sensitivities of 90.8% and 97.9% and specificities of 99.1% and 94.0% for detecting PM and TF, respectively. CONCLUSIONS: The proposed DCNN-DS approach demonstrated reliable performance with high sensitivity, specificity, and AUC to classify different MM levels on fundus photographs sourced from clinics. It can help identify MM automatically among the large myopic groups and show great potential for real-life applications.

17.
Onco Targets Ther ; 14: 3769-3781, 2021.
Article in English | MEDLINE | ID: mdl-34168462

ABSTRACT

BACKGROUND: Melanoma is a prevalent skin cancer with the high rate of metastasis and mortality, affecting the increasing number of people worldwide. Bergamottin (BGM) is a natural furanocoumarin derived from grapefruits and presents the potential anti-cancer activity in several tumor models. However, the role of BGM in the development of melanoma remains unclear. Here, we aimed to explore the effect of BGM on the DNA damage and progression of melanoma. METHODS: The effect of BGM on the melanoma progression was analyzed by CCK-8 assays, colony formation assays, transwell assays, Annexin V-FITC Apoptosis Detection Kit, cell-cycle analysis, in vivo tumorigenicity analysis. The mechanism investigation was performed using luciferase reporter gene assays, qPCR assays, and Western blot analysis. RESULTS: We identified that BGM repressed cell proliferation, migration, and invasion of melanoma cells. BGM induced cell cycle arrest at the G0/G1 phase and enhanced apoptosis of melanoma cells. The DNA damage in the melanoma cells was stimulated by the BGM treatment. Meanwhile, BGM was able to up-regulate the expression of miR-145 and miR-145 targeted Cyclin D1 in the melanoma cells. Furthermore, BGM inhibited the progression of melanoma by targeting miR-145/Cyclin D1 axis in vitro. BGM attenuated the tumor growth of melanoma in vivo. CONCLUSION: Thus, we conclude that BGM induces DNA damage and inhibits tumor progression in melanoma by modulating the miR-145/Cyclin D1 axis. Our finding provides new insights into the mechanism by which BGM modulates the development of melanoma. BGM may be applied as a potential anti-tumor candidate for the clinical treatment of melanoma.

18.
Stem Cells Dev ; 30(13): 683-695, 2021 07 01.
Article in English | MEDLINE | ID: mdl-34030475

ABSTRACT

Homeobox B cluster (HoxB) genes play important roles in retinoic acid (RA)-induced early embryonic stem cells (ESCs) differentiation. Knowledge of regulation network of HoxB is important to further unveil the mechanism of ESCs differentiation. In this study, we identified two enhancers that were activated by RA treatment and 4C data showed long-range interactions between HoxB genes and the two enhancers. CRISPR/Cas9-mediated individual or compound deletion of the two enhancers significantly inhibits HoxB gene expression, and transcriptome analysis revealed that RA-induced early ESCs differentiation was blocked in the enhancer KO cells. We propose new mechanism by which two enhancers regulate HoxB gene expression by different regulation modes during RA-induced early ESCs differentiation through long-range chromatin interactions.


Subject(s)
Chromatin , Tretinoin , Cell Differentiation/genetics , Chromatin/genetics , Chromatin/metabolism , Embryonic Stem Cells , Homeodomain Proteins/genetics , Homeodomain Proteins/metabolism , Tretinoin/metabolism , Tretinoin/pharmacology
19.
Stem Cell Res ; 49: 102097, 2020 12.
Article in English | MEDLINE | ID: mdl-33271468

ABSTRACT

The developmental plasticity of embryonic stem cells (ESCs) is mainly controlled by well-characterized transcription factors, but additional factors, especially those related to metabolism that modulate this intrinsic program remain elusive. Here, using whole transcriptome analysis, we identified branched-chain amino acid aminotransferase-1(Bcat1) as highly-expressed in mouse ESCs and dramatically down-regulated upon differentiation. Bcat1 deletion impaired pluripotency and self-renewal in mouse ESCs, while Bcat1 overexpression resulted in robust ESC self-renewal and inhibition of differentiation. Whole genome bisulfite sequencing (WGBS) analysis showed that Bcat1 deletion altered whole genome methylation levels and hence gene expression in multiple pathways. Specifically, Bcat1 deletion increased expression of RAS protein activator like 1(Rasal1), leading to inactivation of Ras-Erk/MAPK signaling, while Rasal1 inhibition rescued defects seen in Bcat1 deleted cells. In summary, we demonstrate that Bcat1 is essential for mouse ESC self-renewal and pluripotency and that this effect is mediated by DNA methylation and the Ras signaling pathway.


Subject(s)
Embryonic Stem Cells , Mouse Embryonic Stem Cells , Transaminases/genetics , ras Proteins/metabolism , Animals , Cell Differentiation , Mice , Signal Transduction
20.
Int J Oncol ; 54(5): 1567-1578, 2019 May.
Article in English | MEDLINE | ID: mdl-30896790

ABSTRACT

Emerging evidence indicates that mesenchymal stem cells (MSCs) serve an indispensable role in the tumor microenvironment. However, whether MSCs participate in the development of oral carcinogenesis remains unclear. The present study isolated MSCs from clinical tissues and investigated the differences of MSCs derived from normal oral mucosa (N­MSC), oral leukoplakia with dysplasia (LK­MSC) and oral carcinoma (Ca­MSC). The results revealed that the LK­MSCs exhibited reduced proliferation and migration, compared with the N­MSCs and Ca­MSCs. Furthermore, it was demonstrated that the exosomes secreted by LK­MSCs have significant roles in promoting proliferation, migration and invasion in vitro, which was similar to the Ca­MSC­derived exosomes. The promoting effect was also demonstrated in a 3D coculture model. When the secretion of exosomes was blocked, the promoting effect of LK­MSCs was reversed. Based on a microarray analysis of MSC­derived exosomes, microRNA­8485 (miR­8485) was identified to be ectopically expressed. The exosomal miR­8485 was capable of promoting the proliferation, migration and invasion of tumor cells. Therefore, the present study highlights the significance of MSC­derived exosomes and exosomal miR­8485 in premalignant lesions and carcinogenesis. Intervention with the secretion of MSC­derived­exosomes may be an innovative strategy to retard the carcinogenesis.


Subject(s)
Exosomes/pathology , Leukoplakia, Oral/pathology , MicroRNAs/genetics , Mouth Mucosa/cytology , Mouth Neoplasms/pathology , Adult , Aged , Cell Differentiation , Cell Line, Tumor , Cell Movement , Cell Proliferation , Cells, Cultured , Coculture Techniques/methods , Exosomes/genetics , Female , Gene Expression Regulation, Neoplastic , Humans , Leukoplakia, Oral/genetics , Male , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/metabolism , Middle Aged , Mouth Mucosa/metabolism , Mouth Mucosa/pathology , Mouth Neoplasms/genetics , Tumor Microenvironment , Up-Regulation
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