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1.
Mol Cell Biochem ; 478(7): 1465-1473, 2023 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-36370237

RESUMO

LRRC1 is a regulator of cellular polarity that is expressed at high levels in a range of tumor tissue types. Here, we conducted an analysis of the previously unexplored role of LRRC1 as a component of the adipogenic differentiation network. During the early stage (days 3-7) adipocytic differentiation of human mesenchymal stem cells (MSCs), LRRC1 was found to be upregulated at both the mRNA and protein levels. Moreover, the expression of LRRC1 was found to be controlled by PPARγ, which is a key transcriptional regulator of adipogenesis. Inhibiting LRRC1 expression reduced the adipogenic potential of hMSCs, with a concomitant reduction in the expression of three adipogenesis-associated proteins (SCD, LIPE, FASN). Together, these data offer new insight into the functional importance of LRRC1 both in general and in the context of adipocytic differentiation.


Assuntos
Células-Tronco Mesenquimais , Neoplasias , Humanos , PPAR gama/genética , PPAR gama/metabolismo , Células-Tronco Mesenquimais/metabolismo , Diferenciação Celular , Adipogenia/genética , Neoplasias/metabolismo , Células Cultivadas , Proteínas de Transporte/metabolismo , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo
2.
Oral Dis ; 29(8): 3472-3480, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36070195

RESUMO

OBJECTIVES: Hemifacial microsomia (HFM) is a common birth defect involving the first and second branchial arch derivatives. Although several chromosomal abnormalities and causal gene variants have been identified, genetic etiologies in a majority of cases with HFM remain unknown. This study aimed to identify genetic mutations in affected individuals with HFM. METHODS: Whole-exome sequencing and bioinformatics analysis were performed for 16 affected individuals and their family members. Sanger sequencing was applied for confirmation of selected mutations. Zebrafish embryos were used for in situ hybridization of candidate gene, microinjection with antisense morpholino, and cartilage staining. RESULTS: A homozygous missense mutation (c.484G > A; p.V162I) in the FRK gene was identified in an 18-year-old girl with HFM and dental abnormalities. Heterozygous mutation of this mutation was identified in her parents, who are first cousins in a consanguineous family. FRK is highly expressed in the Meckel's cartilage during embryonic development in mouse and zebrafish. Knockdown of frk in zebrafish showed a lower length and width ratio of Meckel's cartilage, abnormal mandibular jaw joint, and disorganized ceratobranchial cartilage and bone. CONCLUSIONS: We identified a recessive variant in the FRK gene as a novel candidate gene for a patient with HFM and mandibular hypoplasia and revealed its effects on craniofacial and embryonic development in zebrafish.


Assuntos
Síndrome de Goldenhar , Humanos , Feminino , Camundongos , Animais , Adolescente , Síndrome de Goldenhar/genética , Peixe-Zebra/genética , Mandíbula/anormalidades , Articulação Temporomandibular , Cartilagem , Proteínas de Neoplasias , Proteínas Tirosina Quinases
3.
Zhong Nan Da Xue Xue Bao Yi Xue Ban ; 44(2): 122-127, 2019 Feb 28.
Artigo em Zh | MEDLINE | ID: mdl-30837379

RESUMO

OBJECTIVE: To clone human mitogen-activated protein kinase kinase 6 (MKK6) gene promoter and explore its transcription activity by ubiquitin specific peptidase 22 (USP22).
 Methods: MKK6 gene promoter was amplified by PCR and two bases mutation within USP22 binding site was subsequently introduced. The wild type and mutant MKK6 promoter were inserted into the luciferase report vector pGL3-Basic, respectively. Recombinant plasmids were co-transfected with plasmid pRL-TK into HeLa cells, and the luciferase activities were measured by dual luciferase reporter system. Furthermore, the direct interaction between USP22 and MKK6 promoter was detected by chromatin immunoprecipitation (ChIP) assay. Finally, the MKK6 transcription activity was measured after knockdown of USP22.
 Results: The recombinant luciferase report vectors containing wild or mutant type of MKK6 promoter were successfully constructed. Mutation of USP22 binding site resulted in decrease of MKK6 promoter-driven luciferase activity in HeLa cells (P<0.05). USP22 could interact directly with MKK6 promoter. Down-regulation of USP22 led to the decreased MKK6 mRNA expression (P<0.05).
 Conclusion: USP22 could regulate the transcription activity of MKK6 gene in HeLa cells.


Assuntos
Tioléster Hidrolases/metabolismo , Células HeLa , Humanos , Luciferases , MAP Quinase Quinase 6 , Regiões Promotoras Genéticas , Transcrição Gênica , Ubiquitina Tiolesterase
4.
Mol Cell Biochem ; 449(1-2): 295-303, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29959592

RESUMO

MicroRNAs are members of the family of non-coding small RNAs that regulate gene expression either by inhibiting mRNA translation or by promoting mRNA degradation at the post-transcriptional level. They play an important role in the differentiation of human bone marrow mesenchymal stem cells (hMSCs) into adipocytes. However, the role of microRNAs in this process remains to be poorly understood. Here, we observed that miR-377-3p expression was markedly decreased during adipogenic differentiation of hMSCs. Overexpression of miR-377-3p decreased adipocyte differentiation and downregulated the expression of adipogenic markers. Meanwhile, bioinformatics-based studies suggested that LIFR is a target of miR-377-3p. Further analysis confirmed that expression of LIFR present markedly increased during adipogenic differentiation of hMSCs. In addition, downregulation expression of LIFR significantly inhibited the process of adipocyte differentiation. To confirm the relation between miR-377-3p and LIFR, luciferase reporter assays were carried out. The results indicated that miR-377-3p bound directly to the 3'-untranslated region of LIFR. These data indicate that miR-377-3p suppressed adipogenesis of hMSCs by targeting LIFR, which provides novel insights into the molecular mechanism of miRNA-mediated cellular differentiation.


Assuntos
Adipogenia , Células da Medula Óssea/metabolismo , Diferenciação Celular , Subunidade alfa de Receptor de Fator Inibidor de Leucemia/biossíntese , Células-Tronco Mesenquimais/metabolismo , MicroRNAs/metabolismo , Regiões 3' não Traduzidas , Células da Medula Óssea/citologia , Linhagem Celular , Humanos , Subunidade alfa de Receptor de Fator Inibidor de Leucemia/genética , Células-Tronco Mesenquimais/citologia , MicroRNAs/genética
5.
Genet Mol Biol ; 41(2): 488-495, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30088609

RESUMO

The ubiquitin-specific protease 22 (USP22) is an oncogene and its expression is upregulated in many types of cancer. In the nucleus, USP22 functions as one subunit of the SAGA to regulate gene transcription. However, the genome-wide USP22 binding sites and its direct target genes are yet clear. In this study, we characterized the potential genomic binding sites of UPS22 and GCN5 by ChIP-seq using specific antibodies in HeLa cells. There were 408 overlapping putative target genes bound by both USP22 and GCN5. Motif analysis showed that the sequences bound by USP22 and GCN5 shared two common motifs. Gene ontology (GO) and pathway analysis indicated that the genes targeted by USP22 and GCN5 were involved in different physiological processes and pathways. Further RNA-seq, GO and pathway analyses revealed that knockdown of UPS22 induced differential expression of many genes that participated in diverse physiological processes, such as metabolic process. Integration of ChIP-seq and RNA-seq data revealed that UPS22 bound to the promoters of 56 genes. These findings may provide new insights into the regulation of USP22 on gene expression during the development of cervical cancer.

6.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 48(6): 824-827, 2017 Nov.
Artigo em Zh | MEDLINE | ID: mdl-29260514

RESUMO

OBJECTIVE: To construct the expression vectors for red fluorescent protein fused with inhibiting peptides for nuclear import (Bimax),and explore the location of Bimax and its potential effects on cell proliferation and migration in HeLa cells. METHODS: Two kinds of polynucleotide encoding inhibiting peptides for nuclear import were synthesis respectively and subsequently annealed for inserting into vector pDs-Red-C1. The recombinant plasmids were transfected into competent bacterial DH-5α. After transfection,the positive bacteria were picked up for DNA sequencing. The recombinant plasmids pDs-Red-Bimax2,pDs-Red-Bimax1 and negative plasmid pDs-Red-C1 were transfected into HeLa cells respectively according to Lipofectamine2000 protocol. After transfection,the expression and location of red fluorescent protein were observed with fluorescence microscope. Furthermore,MTT assay and cell-migration assay were used to detect the proliferation and migration of Bimax transducted cells. RESULTS: DNA sequencing showed that the polynucleotides encoding Bimax1 or Bimax2 were inserted into pDs-Red-C1 vector successfully. After transfected into HeLa cells,the inhibiting peptide induced red fluorescent protein locating in nuclear. Furthermore,either the fusion protein RFP-Bimax1 or RFP-Bimax2 can suppress the proliferation and migration of HeLa cells. CONCLUSION: The expression vectors for red fluorescent protein fused with inhibiting peptides for nuclear import were successfully constructed. In addition,the fusion proteins were expressed and located in nuclear and suppressed the proliferation and migration of tumor cells.


Assuntos
Transporte Ativo do Núcleo Celular , Vetores Genéticos , Peptídeos , Plasmídeos , Movimento Celular , Células HeLa , Humanos , Proteínas Luminescentes , Transfecção , Proteína Vermelha Fluorescente
7.
Biochem Biophys Res Commun ; 449(1): 14-8, 2014 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-24802393

RESUMO

Ubiquitin-specific processing enzyme 22 (USP22), a member of the deubiquitinase family, is over-expressed in most human cancers and has been implicated in tumorigenesis. Because it is an enzymatic subunit of the human SAGA transcriptional cofactor, USP22 deubiquitylates histone H2A and H2B in the nucleus, thus participating in gene regulation and cell-cycle progression. However, the mechanisms regulating its nuclear translocation have not yet been elucidated. It was here demonstrated that USP22 is imported into the nucleus through a mechanism mediated by nuclear localization signal (NLS). The bipartite NLS sequence KRELELLKHNPKRRKIT (aa152-168), was identified as the functional NLS for its nuclear localization. Furthermore, a short cluster of basic amino acid residues KRRK within this bipartite NLS plays the primary role in nuclear localization and is evolutionarily conserved in USP22 homologues. In the present study, a functional NLS and the minimal sequences required for the active targeting of USP22 to the nucleus were identified. These findings may provide a molecular basis for the mechanism underlying USP22 nuclear trafficking and function.


Assuntos
Núcleo Celular/química , Núcleo Celular/fisiologia , Sinais de Localização Nuclear/química , Sinais de Localização Nuclear/fisiologia , Tioléster Hidrolases/química , Tioléster Hidrolases/fisiologia , Transporte Ativo do Núcleo Celular/fisiologia , Sequência de Aminoácidos , Sequência Conservada , Células HeLa , Humanos , Dados de Sequência Molecular , Relação Estrutura-Atividade , Ubiquitina Tiolesterase
8.
Zhong Nan Da Xue Xue Bao Yi Xue Ban ; 39(8): 764-8, 2014 Aug.
Artigo em Zh | MEDLINE | ID: mdl-25202959

RESUMO

OBJECTIVE: To clone 5' untranslated region of human IPO8 gene and determine its transcription activity. METHODS: We used 5' rapid amplification of cDNA ends (RACE) analysis to identify the IPO8 transcription start site (TSS), and amplified series truncated 5' UTR fragment containing transcription start site. The PCR productions were inserted into luciferase report vector pGL3- Basic. After confirmation by restriction enzyme digestion, the recombinant plasmids were cotransfected into Saos-2 cells with plasmid pRL-TK. The luciferase activities were measured by dual luciferase reporter system. RESULTS: The IPO8 gene transcription start site was established. The electrophoresis analysis of restriction enzyme digestion and DNA sequencing verified the fragments were successfully amplified and inserted into pGL3-Basic. After the recombinant plasmids transfected, the highexpressions of luciferase were detected in Saos-2 cells. CONCLUSION: The recombinant vector containing IPO8 promoter is constructed successfully, which provides a foundation for determining expressional regulation of IPO8 in the further study.


Assuntos
Regiões Promotoras Genéticas , beta Carioferinas/genética , Clonagem Molecular , DNA Complementar , Vetores Genéticos , Humanos , Luciferases , Plasmídeos , Transfecção
9.
Head Neck ; 46(3): 513-527, 2024 03.
Artigo em Inglês | MEDLINE | ID: mdl-38108536

RESUMO

BACKGROUND: The purpose of this study was to explore preliminary the performance of radiomics machine learning models based on multimodal MRI to predict the risk of cervical lymph node metastasis (CLNM) for oral tongue squamous cell carcinoma (OTSCC) patients. METHODS: A total of 400 patients were enrolled in this study and divided into six groups according to the different combinations of MRI sequences. Group I consisted of patients with T1-weighted images (T1WI) and FS-T2WI (fat-suppressed T2-weighted images), group II consisted of patients with T1WI, FS-T2WI, and contrast enhanced MRI (CE-MRI), group III consisted of patients with T1WI, FS-T2WI, and T2-weighted images (T2WI), group IV consisted of patients with T1WI, FS-T2WI, CE-MRI, and T2WI, group V consisted of patients with T1WI, FS-T2WI, T2WI, and apparent diffusion coefficient map (ADC), and group VI consisted of patients with T1WI, FS-T2WI, CE-MRI, T2WI, and ADC. Machine learning models were constructed. The performance of the models was compared in each group. RESULTS: The machine learning model in group IV including T1WI, FS-T2WI, T2WI, and CE-MRI presented best prediction performance, with AUCs of 0.881 and 0.868 in the two sets. The models with CE-MRI performed better than the models without CE-MRI(I vs. II, III vs. IV, V vs. VI). CONCLUSIONS: The radiomics machine learning models based on CE-MRI showed great accuracy and stability in predicting the risk of CLNM for OTSCC patients.


Assuntos
Carcinoma de Células Escamosas , Neoplasias de Cabeça e Pescoço , Neoplasias da Língua , Humanos , Metástase Linfática , Carcinoma de Células Escamosas de Cabeça e Pescoço , Carcinoma de Células Escamosas/diagnóstico por imagem , Radiômica , Neoplasias da Língua/diagnóstico por imagem , Imageamento por Ressonância Magnética/métodos , Aprendizado de Máquina , Estudos Retrospectivos
10.
Adv Mater ; 36(5): e2307817, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-37948543

RESUMO

Advanced chemotherapeutic strategies including prodrug and nanocatalytic medicine have significantly advanced tumor-selective theranostics, but delicate prodrug screening, tedious synthesis, low degradability/biocompatibility of inorganic components, and unsatisfied reaction activity complicate treatment efficacies. Here, the intrinsic anticancer bioactivity of liquid metal nanodroplets (LMNDs) is explored through galvanic replacement. By utilizing a mechano-degradable ligand, the resultant size of the aqueous LMND is unexpectedly controlled as small as ≈20 nm (LMND20). It is demonstrated that LMND20 presents excellent tumor penetration and biocompatibility and activates tumor-selective carrier-to-drug conversion, synchronously depleting Cu2+ ions and producing Ga3+ ions through galvanic replacement. Together with abundant generation of reactive oxygen species, multiple anticancer pathways lead to selective apoptosis and anti-angiogenesis of breast cancer cells. Compared to the preclinical/clinical anticancer drugs of tetrathiomolybdate and Ga(NO3 )3 , LMND20 administration significantly improves the therapeutic efficacy and survival in a BCap-37 xenograft mouse model, yet without obvious side effects.


Assuntos
Antineoplásicos , Neoplasias , Pró-Fármacos , Humanos , Animais , Camundongos , Nanomedicina , Neoplasias/tratamento farmacológico , Neoplasias/metabolismo , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Pró-Fármacos/farmacologia , Pró-Fármacos/uso terapêutico , Metais , Íons , Linhagem Celular Tumoral
11.
Aging (Albany NY) ; 15(15): 7637-7654, 2023 08 04.
Artigo em Inglês | MEDLINE | ID: mdl-37543430

RESUMO

The delicate equilibrium between osteoblast and adipocyte differentiation of MSCs is highly regulated. We screened for early-stage osteogenesis- or adipogenesis-based MSCs protein expression profiles using TMT-based quantitative proteomic analysis to identify novel participating molecules. Protein annotation, hierarchical clustering, functional stratification, and protein-protein association assessments were performed. Moreover, two upregulated proteins, namely, FBLN2 and NPR3, were validated to participate in the osteogenic differentiation process of MSCs. After that, we independently downregulated FBLN2 and NPR3 over seven days of osteogenic differentiation, and we performed quantitative proteomics analysis to determine how different proteins were regulated in knockdown vs. control cells. Based on gene ontology (GO) and network analyses, FBLN2 deficiency induced functional alterations associated with biological regulation and stimulus-response, whereas NPR3 deficiency induced functional alterations related to cellular and metabolic processes, and so on. These findings suggested that proteomics remains a useful method for an in-depth study of the MSCs differentiation process. This will assist in comprehensively evaluating its role in osteoporosis and provide additional approaches for identifying as-yet-unidentified effector molecules.


Assuntos
Células-Tronco Mesenquimais , Osteogênese , Osteogênese/genética , Proteômica , Diferenciação Celular/fisiologia , Adipogenia , Células-Tronco Mesenquimais/metabolismo
12.
J Craniomaxillofac Surg ; 51(9): 574-579, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37604768

RESUMO

The purpose of this study was to introduce a novel individualized flap design method for large anterior floor of the mouth (AFOM) defect reconstruction, review experience with the use of this flap design method for large AFOM defect reconstruction, and assess its functional results. A retrospective study of patients who received large AFOM defect reconstruction with free flaps was conducted. There was a cohort of patients who were treated using the novel individualized flap design method and a cohort without flap design. Functional outcomes were evaluated with appropriate scales. Outcomes were analyzed, and a p-value <0.05 was considered significant. 22 patients received the individualized flap design, while 21 patients were treated without a special flap design. All flaps survived. All free flaps harvested with the novel individualized flap design method better matched AFOM defects. Relative to patients without flap design, patients in the novel individualized flap design group showed significant improvement in speech intelligibility (p = 0.036) and swallowing function (p = 0.019). Within the limitation of the study it seems that large AFOM defect reconstruction with the novel individualized flap design method can not only cover and close the wound to avoid oral-neck fistulae, but also maintains tongue mobility to achieve better functional outcomes than in patients without flap design.

13.
Oral Oncol ; 145: 106531, 2023 10.
Artigo em Inglês | MEDLINE | ID: mdl-37499327

RESUMO

BACKGROUND: To achieve improved functional outcomes in subtotal tongue reconstruction, a flap design with sufficient volume and appropriate shape is necessary. In this study, we introduce an "Individualized and Convenient Tongue Model" (ICTM) for flap design in subtotal tongue reconstruction. METHODS: By studying the anatomical morphology of the tongue, we found a similar geometry within the dorsum and body of the tongue as well as the mouth floor. This can be used to create an ICTM through folding and splicing. We can simulate tongue defects in the ICTM and transform defect shapes into guide plates for flap design. In this study, fifty-eight patients requiring subtotal tongue reconstruction were randomly divided into two groups: an ICTM group (35 patients) and a conventional group (31 patients). In the ICTM group, we individually designed profunda artery perforator flaps (PAPFs) or anterolateral thigh flaps (ALTFs) using the ICTM method. In the conventional group, the flap was designed according to the surgeon's clinical experience. Patient demographics, operative and follow-up data were recorded. Swallowing, speech intelligibility, and cosmetic results were assessed using appropriate scales. RESULTS: All flaps survived, although there were no significant differences in tumor size, operation time, flap size, and complication rate compared to the conventional group. Patients in the ICTM group had significantly improved speech intelligibility (p = 0.019), cosmetic appearance (p = 0.009), and swallowing ability (p = 0.003). CONCLUSIONS: The ICTM technique is an effective and convenient solution for subtotal tongue reconstruction that provides an individualized flap design and improves functional outcomes compared to the conventional design.


Assuntos
Retalho Perfurante , Procedimentos de Cirurgia Plástica , Neoplasias da Língua , Humanos , Neoplasias da Língua/cirurgia , Neoplasias da Língua/patologia , Língua/cirurgia , Língua/patologia , Retalho Perfurante/cirurgia , Soalho Bucal/patologia
14.
Front Vet Sci ; 10: 1237780, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37781284

RESUMO

The significant morphological differences and abundant germplasm resources of Chinese indigenous dog breeds can be attributed to the diverse geographical environment, including plateaus, mountains, and a long history of raising dogs. The combination of both natural and artificial selection during the past several thousand years has led to hundreds of dog breeds with distinct morphological traits and environmental adaptations. China is one of the earliest countries to domesticate dogs and there are more than 50 ancient indigenous dog breeds. In this study, the run of homozygosity (ROH) and proportion of the autosomal genome covered by ROHs (FROH) were calculated for 10 dog breeds that are the most representative Chinese indigenous dogs based on 170K SNP microarray. The results of FROH showed that the Chuandong hound dogs (HCSSC) have the highest level of inbreeding among the tested breeds. The inbreeding in HCSSC occurred more recently than the Liangshan dogs (SCLSQ) dogs because of more numbers of long ROHs in HCSSC dogs, and the former also have higher inbreeding degree. In addition, there are significant differences in the inbreeding degree among different subpopulations of the same breed, such as the Thin dogs from Shaanxi and Shandong province. To explore genome-wide selection signatures among different breeds, including coat color, ear shape, and altitude adaptability, we performed genome selection analyses of FST and cross population extended haplotype homozygosity (XP-EHH). For the coat color, the FST analysis between Xiasi dogs (XSGZ) and HCSSC dogs was performed and identified multiple genes involved in coat color, hair follicle, and bone development, including MC1R, KITLG, SOX5, RSPO2, and TBX15. For the plateau adaptability, we performed FST and XP-EHH analyses between dogs from Tibet (Tibetan Mastiffs and Nyingchi dogs) and plain regions (Guangxi Biwei dogs GXBWQ and Guandong Sharpei dogs). The results showed the EPAS1 gene in dogs from Tibet undergo strong selection. Multiple genes identified for selection signals based on different usage of dogs. Furthermore, the results of ear shape analyses showed that MSRB3 was likely to be the main gene causing the drop ear of domestic dogs. Our study provides new insights into further understanding of Chinese indigenous dogs.

15.
Folia Histochem Cytobiol ; 60(2): 179-190, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35575247

RESUMO

INTRODUCTION: In osteoporosis field, it had been clinically well established a given relationship between bone formation and lipid accumulation. Although numerous molecules had been well documented for adipogenesis and osteoblastogenesis (adipo-osteoblastogenesis), the reciprocal transcriptional regulation still remains to be explored. MATERIAL AND METHODS: Here, we tried to identify the common candidate genes of adipocyte/osteoblastocyte differentiation at 3, 5, and 7 days using human mesenchymal stem cells (hMSCs) via RNA-Seq technique. By using RNA interference (RNAi), we further confirmed the function of candidate genes during adipo-osteoblastogenesis through Oil Red/Alizarin Red/alkaline phosphatase (ALPL) staining and qRT-PCR (quantitative real-time PCR). RESULTS: The identified 275 significantly differentially expressed genes (DEGs), especially with the down-regulated genes most prevalent and PI3K-AKT signaling pathway mostly enriched, were simultaneously shared by both differentiation events. Using lentiviral system, we further confirmed that ANKRD1 (ankyrin repeat domain 1) promoted adipogenesis and inhibited osteoblastogenesis via RNA interference (RNAi), and IGF1 (insulin like growth factor 1) simultaneously facilitated adipo-osteoblastogenesis on the base of gene expression of biomarkers and cellular phenotype property. CONCLUSION: This study would provide the potential molecular switches to control the adipocyte/osteoblastocyte balance or hMSCs fate choices and clues to screen the study and therapy targets of metabolic bone disease osteoporosis.


Assuntos
Células-Tronco Mesenquimais , Osteoporose , Adipogenia/genética , Diferenciação Celular , Células Cultivadas , Humanos , Células-Tronco Mesenquimais/metabolismo , Osteoblastos/metabolismo , Osteogênese , Osteoporose/genética , Osteoporose/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo
16.
BMC Med Genomics ; 15(1): 17, 2022 01 31.
Artigo em Inglês | MEDLINE | ID: mdl-35101056

RESUMO

Although differential expression of genes is apparent during the adipogenic/osteogenic differentiation of marrow mesenchymal stem cells (MSCs), it is not known whether this is associated with changes in chromosomal structure. In this study, we used ATAC-sequencing technology to observe variations in chromatin assembly during the early stages of MSC differentiation. This showed significant changes in the number and distribution of chromosome accessibility at different time points of adipogenic/osteogenic differentiation. Sequencing of differential peaks indicated alterations in transcription factor motifs involved in MSC differentiation. Gene Ontology (GO) and pathway analysis indicated that changes in biological function resulted from the alterations in chromatin accessibility. We then integrated ATAC-seq and RNA-seq and found that only a small proportion of the overlapping genes were screened out from ATAC-seq and RNA-seq overlapping. Through GO and pathway analysis of these overlapped genes, we not only observed some known biological functions related to adipogenic/osteogenic differentiation but also noticed some unusual biological clustering during MSC differentiation. In summary, our work not only presents the landscape of chromatin accessibility of MSC during differentiation but also helps to further our understanding of the underlying mechanisms of gene expression in these processes.


Assuntos
Cromatina , Células-Tronco Mesenquimais , Adipócitos/metabolismo , Diferenciação Celular/genética , Cromatina/genética , Cromatina/metabolismo , Humanos , Células-Tronco Mesenquimais/metabolismo , Osteoblastos/metabolismo , Osteogênese
17.
Front Genet ; 13: 1027396, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36468022

RESUMO

Background: Hearing loss (HL) is the most common form of sensory disorder in humans. Molecular diagnosis of HL is important for genetic counseling for the affected individuals and their families. Methods: To identify potential genetic causes, we performed whole-exome sequencing and related biomedical informatics for 351 non-syndromic HL patients and their family members. Results: In the present study, we report the identification of four compound heterozygous variants in the CDH23 gene from four affected families, including four novel variants (c.995C>A, p.T332K; c.2159G>A, p.R720Q; c.5534A>G, p.N1845S, and c.7055-1G>C) and two frequently reported variants (c.719C>T, p.P240L and c.4762C>T, p.R1588W). Conclusion: Our findings significantly expanded the mutation spectrum of CDH23-associated autosomal recessive hearing loss.

18.
Folia Histochem Cytobiol ; 60(1): 89-100, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35212388

RESUMO

INTRODUCTION: With cellular lipid storage varying, the balance between lipid intake and lipid degradation was a must to keep healthy and determined the level of lipid droplets. Although lipid droplets accumulation had been well demonstrated in adipocytes, gene expression profiling and gene function during adipogenesis and osteoblastogenesis remain unknown. MATERIAL AND METHODS: Here, this work profiled gene transcriptional landscapes of lipid droplets formation during adipogenesis from human mesenchymal stem cells (hMSCs) using RNA-Seq technique. By using RNA interference (RNAi) we investigated the function of candidate genes during adipogenesis and osteoblastogenesis using Oil Red/Alizarin Red/alkaline phosphatase (ALPL) staining and qRT-PCR (quantitative real-time PCR). RESULTS: Eleven differentially up-regulated genes associated with lipid droplets formation were identified at 3, 5, 7, 14, 21, and 28 days during adipogenesis. Unexpectedly, APOB per se inhibiting adipogenesis weakened osteoblastogenesis and METTL7A facilitating adipogenesis negligibly inhibited osteoblastogenesis according to the phenotypic characterization of adipocytes and osteoblasts and transcriptional condition of biomarkers through lentivirus transfection assays. CONCLUSIONS: The establishment of the gene transcriptional profiling of lipid droplets formation would provide the molecular switches of hMSCs cell fate determination and the study targets for fat metabolic diseases.


Assuntos
Adipogenia , Células-Tronco Mesenquimais , Adipócitos , Adipogenia/genética , Diferenciação Celular , Células Cultivadas , Humanos , Gotículas Lipídicas , Células-Tronco Mesenquimais/metabolismo
19.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 42(2): 166-9, 2011 Mar.
Artigo em Zh | MEDLINE | ID: mdl-21500546

RESUMO

OBJECTIVE: To construct eukaryotic expression plasmid expressing siRNA targeting ubiquitin specific peptidase 22 gene (USP22), and to investigate its effect on the growth of hepatoma carcinoma cells HepG2. METHODS: siRNA templates were synthesized based on USP22 mRNA sequence and cloned into vector Pmscv/Hyg/U6. The resulting recombinant was identified by restriction enzyme digestion and DNA sequencing. Recombinants were than transfected into HepG2 cells mediated by liposome. The USP22 protein and mRNA in HepG2 cells were detected by western blot and RT-PCR, respectively. The cellular growth activity was evaluated with MTT assay. RESULTS: Recombinant plasmid expressing siRNA targeting USP22 was successfully constructed. The down-regulated protein and mRNA level of USP22 and decreased cellular growth in HepG2 cells transfected with recombinant plasmid were observed. CONCLUSION: The eukaryotic expression vector for RNA interference USP22 gene is constructed successfully, which inhibits the expression of USP22 in HepG2 cells and suppresses cell proliferation.


Assuntos
Vetores Genéticos/genética , Interferência de RNA , RNA Interferente Pequeno/genética , Tioléster Hidrolases/genética , Sequência de Bases , Proliferação de Células , Células Hep G2 , Humanos , Dados de Sequência Molecular , Plasmídeos/genética , RNA Mensageiro/genética , Transfecção , Ubiquitina Tiolesterase
20.
Front Genet ; 12: 728020, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34912366

RESUMO

A cohort of 542 individuals in 166 families with congenital hearing loss was recruited for whole-exome sequencing analysis. Here, we report the identification of three variants in five affected individuals in two unrelated families. In family 1, a nonsense mutation (c.1516C>T, p.R506*) in the ATP6V1B2 gene, a known causal allele for dominant deafness-onychodystrophy (DDOD), was identified in the mother and son with DDOD. However, a novel heterozygous variant (c.1590T>G, p.D530E) in TJP2, a known causal gene for hearing-loss, was also detected in the patients. In family 2, the same mutation (c.1516C>T, p.R506*) of ATP6V1B2 was detected from the father and daughter with DDOD. Furthermore, a novel heterozygous variant (c.733A>G, p.M245V) in the KIF11 gene was identified from the spouse with sensorineural hearing-loss and epilepsy. Notably, genotype-phenotype analysis of KIF11-associated disorders revealed that the p.M245V and two reported hearing-loss-associated variants (p.S235C and p.H244Y) are all mapped to a single ß-sheet (Ser235∼M245) in the kinesin motor domain. Together, this is the first demonstration that ATP6V1B2-caused DDOD is an autosomal dominant genetic disease, compared to previous cases with de novo mutation. Our findings expand the variant spectrum of hearing-loss-associated genes and provide new insights on understanding of hearing-loss candidate genes ATP6V1B2, TJP2, and KIF11.

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