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1.
Adv Exp Med Biol ; 1287: 1-7, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33034022

RESUMO

The evolutionary conserved Notch pathway that first developed in metazoans and that was first discovered in fruit flies (Drosophila melanogaster) governs fundamental cell fate decisions and many other cellular key processes not only in embryonic development but also during initiation, promotion, and progression of cancer. On a first look, the Notch pathway appears remarkably simple, with its key feature representing a direct connection between an extracellular signal and transcriptional output without the need of a long chain of protein intermediaries as known from many other signaling pathways. However, on a second, closer look, this obvious simplicity exerts surprising complexity. There is no doubt that the enormous scientific progress in unraveling the functional mechanisms that underlie this complexity has recently greatly increased our knowledge about the role of Notch signaling for pathogenesis and progression of many types of cancer. Moreover, these new scientific findings have shown promise in opening new avenues for cancer prevention and therapy, although this goal is still challenging. Vol. III of the second edition of the book Notch Signaling in Embryology and Cancer, entitled Notch Signaling in Cancer, summarizes important recent developments in this fast-moving and fascinating field. Here, we give an introduction to this book and a short summary of the individual chapters that are written by leading scientists, covering the latest developments in this intriguing research area.


Assuntos
Neoplasias/prevenção & controle , Neoplasias/terapia , Receptores Notch/metabolismo , Transdução de Sinais , Animais , Diferenciação Celular , Humanos , Neoplasias/patologia
2.
Shanghai Kou Qiang Yi Xue ; 29(3): 225-230, 2020 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-33043336

RESUMO

PURPOSE: To investigate the effects of exendin-4(EX-4) on proliferation, migration and osteogenic differentiation of human periodontal ligament stem cells(PDLSCs). METHODS: PDLSCs were isolated and cultured using limited dilution method in vitro. Colony formation assay, osteogenic and adipogenic differentiation were applied to identify the stem cells. Immunofluorescence staining was used to detect the expression of EX-4 receptor glucagon-like peptide-1 receptor (GLP-1R) on the surface of PDLSCs. PDLSCs were stimulated with 5, 10, 20 or 50 nmol/L EX-4 in vitro. CCK-8, Transwell assay and alkaline phosphatase(ALP) activity assay were used to determine the effects of EX-4 on PDLSCs proliferation, migration and osteogenic differentiation. Quantitative real-time polymerase chain reaction was used to determine the expression of osteogenic related genes ALP, runt-related transcription factor 2(Runx2) and osteocalcin (OCN). The data were analyzed by Graphpad Prims 6.0 software package. RESULTS: PDLSCs were successfully isolated and cultivated. GLP-1R positively expressed on the surface of PDLSCs. EX-4 exerted no significant effect on PDLSCs proliferation(P>0.05). EX-4 significantly promoted migration, ALP activity and osteogenic related genes expression of PDLSCs (P<0.05). CONCLUSIONS: 10 nmol/L EX-4 could promote migration and osteogenic differentiation of PDLSCs.


Assuntos
Exenatida , Ligamento Periodontal , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Exenatida/farmacologia , Humanos , Osteogênese , Células-Tronco
3.
Shanghai Kou Qiang Yi Xue ; 29(3): 242-249, 2020 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-33043339

RESUMO

PURPOSE: This study was aimed to investigate the mesenchymal stem cell (MSC) phenotypes of immortalized odontoblasts(iODs) and bone morphogenetic protein -2, -4, -6, -7, and -9 (BMPs) differentially regulate the mineralization of iODs. METHODS: ODs were immortalized by SV40 T antigen to establish iOD lineages, and the endogenous expression of BMPs was successively examined. Recombinant adenoviruses expressing BMPs and GFP were generated using Ad-Easy technology. The proliferation capability of iODs was examined using an MTT kit. MSC markers of iODs were examined by immunofluorescence. In vitro, semiquantitative RT-PCR, alkaline phosphatase(ALP) activity assay, matrix mineralization assay and oil red O staining assay were used to examine the osteogenic and adipogenic differentiation capabilities of iODs. Statistical significance among groups was analyzed by one-way analysis of variance and Scheffe's multiple comparison test was SPSS 21.0 software package. Finally, the volume and density of ectopic mineralized tissues formed in vivo were assessed by micro-CT and histological analysis. RESULTS: ODs can be efficiently immortalized by SV40 T antigen, and the resulting iODs maintained an excellent proliferative activity, expressed certain MSC markers and possessed multiple differentiation capabilities. BMP-2 and BMP-9 regulated iODs osteogenic differentiation better than BMP-4, -6, and -7. CONCLUSIONS: Our findings suggest that ODs and osteogenic growth factors such as BMP-2 and BMP-9 can be used as an efficacious strategy for bone tissue engineering.


Assuntos
Células-Tronco Mesenquimais , Osteogênese , Proteínas Morfogenéticas Ósseas , Diferenciação Celular , Odontoblastos
4.
Shanghai Kou Qiang Yi Xue ; 29(3): 257-261, 2020 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-33043341

RESUMO

PURPOSE: To preliminarily explore the cytotoxicological responses of keratinocytes derived from human embryonic stem cells(K-hESCs) to drugs, and to provide a basis for the establishment of a new biosafety evaluation model. METHODS: CCK-8 assay was used to detect cell viability and cytotoxicity. The detection of pharmacological response was observed and compared when K-hESCs directly derived from human embryonic stem cells, human gingival epithelial cells (HGECs), and human immortalized oral epithelial cells (HIOECs) were treated with retinoic acid (RA), 5-fluorouracil(5-FU), dexamethasone(DEX), and penicillin G(PG). Statistical analysis was performed using SPSS 16.0 software package. RESULTS: After drugs were applied to HGECs, HIOEC and K-hESCs, the half inhibitory concentrations (IC50) of RA was 6.1±0.03, 5.62±0.05 and 6.58±0.02, respectively. The IC50 of 5-FU was 1.65±0.02,3.00±0.02 and 1.72±0.04, respectively. The IC50 of DEX was 113.67±0.014,328±0.002 and 126.17±0.05, respectively. The IC50 of PG was 2200±1.34,3795±2.42 and 2880±1.5, respectively. The IC50 of the four drugs between HIOEC and HGECs had significant differences(P<0.05), but there was no significant difference in IC50 between K-hESCs and HGECs(P>0.05). The IC50 of K-hESCs and HIOEC also had significant differences(P<0.05). CONCLUSIONS: IC50 of K-hESCs was closer to HGECs than HIOEC. It was speculated that K-hESCs could simulate the response of normal human cells in cytotoxicity study.


Assuntos
Células-Tronco Embrionárias , Células-Tronco Embrionárias Humanas , Diferenciação Celular , Células Epiteliais , Humanos , Queratinócitos
5.
Zhong Nan Da Xue Xue Bao Yi Xue Ban ; 45(8): 901-908, 2020 Aug 28.
Artigo em Inglês, Chinês | MEDLINE | ID: mdl-33053530

RESUMO

OBJECTIVES: To compare the effect on proliferation of osteoblasts MC3T3-E1 between the concentrated growth factor extract (CGFe) and the platelet-rich fibrin extract (PRFe). METHODS: CGFe and PRFe were prepared. MC3T3-E1 was cultured in DMEM medium containing CGFe (10%, 20%, or 30%) and PRFe (10%, 20%, or 30%). The proliferation of MC3T3-E1 was detected by MTT assay at Day 1, 3, 5, and 7. ALP activity was detected by alkaline phosphatase (ALP) staining at Day 1, 3, 5, and 7, and mRNA expressions of Runt-related transcription factor 2 (Runx2) and Osterix (Osx) were detected by quantitative RT-PCR (RT-qPCR) at Day 3 and 7. RESULTS: Compared with the control group, CGFe and PRFe promoted the proliferation of MC3T3-E1 at Day 1, 3, 5, and 7 (all P<0.05). Except for the first day, the proliferation activity in the CGFe group was higher than that in the PRFe group (all P<0.05). At Day 1, 3, 5, and 7, compared with the control group, the ALP activities in the CGFe group and the PRFe group were significantly increased (all P<0.05). Except for the first day, the ALP activity in the CGFe group was higher than that in the PRFe group (all P<0.05). At Day 3 and 7, compared with the control group, the mRNA expression levels of Osx and Runx2 in the CGFe group and the PRFe group were significantly increased (all P<0.05); compared with PRFe group, the mRNA expression level of Osx in the CGFe group was significantly higher than that in the PRFe group, and the mRNA expression level of Runx2 was significantly lower than that in the PRFe group (all P<0.05). CONCLUSIONS: CGFe could promote the proliferation of MC3T3-E1 stronger than PRFe, which might be related to the increase of ALP activity and up-regulation of Osx expression.


Assuntos
Fibrina Rica em Plaquetas , Diferenciação Celular , Linhagem Celular , Proliferação de Células , Osteoblastos , Extratos Vegetais
6.
Nat Commun ; 11(1): 4964, 2020 10 02.
Artigo em Inglês | MEDLINE | ID: mdl-33009394

RESUMO

Thrombosis leads to platelet activation and subsequent degradation; therefore, replenishment of platelets from hematopoietic stem/progenitor cells (HSPCs) is needed to maintain the physiological level of circulating platelets. Platelet-derived microparticles (PMPs) are protein- and RNA-containing vesicles released from activated platelets. We hypothesized that factors carried by PMPs might influence the production of platelets from HSPCs, in a positive feedback fashion. Here we show that, during mouse acute liver injury, the density of megakaryocyte in the bone marrow increases following an increase in circulating PMPs, but without thrombopoietin (TPO) upregulation. In vitro, PMPs are internalized by HSPCs and drive them toward a megakaryocytic fate. Mechanistically, miR-1915-3p, a miRNA highly enriched in PMPs, is transported to target cells and suppresses the expression levels of Rho GTPase family member B, thereby inducing megakaryopoiesis. In addition, direct injection of PMPs into irradiated mice increases the number of megakaryocytes and platelets without affecting TPO levels. In conclusion, our data reveal that PMPs have a role in promoting megakaryocytic differentiation and platelet production.


Assuntos
Plaquetas/metabolismo , Diferenciação Celular , Micropartículas Derivadas de Células/metabolismo , Megacariócitos/citologia , MicroRNAs/metabolismo , Animais , Sequência de Bases , Linhagem Celular , Endocitose , Perfilação da Expressão Gênica , Humanos , Fígado/lesões , Fígado/patologia , Masculino , Megacariócitos/metabolismo , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , MicroRNAs/genética , Poliploidia , Proteína rhoB de Ligação ao GTP/metabolismo
7.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi ; 34(10): 1313-1321, 2020 Oct 15.
Artigo em Chinês | MEDLINE | ID: mdl-33063499

RESUMO

Objective: To explore the role of joint regulation of Wnt and bone morphogenetic protein (BMP) signaling pathways in the differentiation of human induced pluripotent stem cells (hiPSCs) into cardiomyocytes. Methods: HiPSCs were cultured and observed under inverted phase contrast microscope. Immunofluorescence staining was used to observe the expressions of hiPSCs pluripotent markers (OCT3/4, NANOG, and TRA-1-60). HiPSCs were passaged which were taken for subsequent experiments within the 35th passage. When the fusion degree of hiPSCs was close to 100%, the CHIR99021 (Wnt pathway activator) was added on the 0th day of differentiation. Different concentrations of IWP4 (inhibitor of Wnt production) were added on the 3rd day of differentiation, and the best concentration of IWP4 was added at different time points. The optimal concentration and the best effective period of IWP4 were obtained by detecting the expression of troponin T (TNNT2) mRNA by real-time fluorescence quantitative PCR. Then, on the basis of adding CHIR99021 and IWP4, different concentrations of BMP-4 were added on the 5th day of differentiation, and the best concentration of BMP-4 was added at different time points. The optimal concentration and best effective period of BMP-4 were obtained by detecting the expression of TNNT2 mRNA. Finally, hiPSCs were divided into three groups: Wnt group, BMP group, and Wnt+BMP group. On the basis of adding CHIR99021 on the 0th day of differentiation, IWP4, BMP-4, and IWP4+BMP-4 were added into Wnt group, BMP group, and Wnt+BMP group respectively according to the screening results. Cells were collected on the 7th and the 15th days of differentiation. The expressions of myocardial precursor cell markers [ISL LIM homeobox 1 (ISL1), NK2 homeobox 5 (NKX2-5)] and cardiomyocyte specific markers [myocyte enhancer factor 2C (MEF2C), myosin light chain 2 (MYL2), MYL7, and TNNT2] were detected by real-time fluorescent quantitative PCR. Cells were collected on the 28th day of differentiation, and the expression of cardiac troponin T (cTnT) was detected by flow cytometry and immunofluorescence staining. Results: The results of cell mophology and immunoflurescence staining showed that the OCT3/4, NANOG, and TRA-1-60 were highly expressed in hiPSCs, which suggested that hiPSCs had characteristics of pluripotency. The optimal concentration of IWP4 was 10.0 µmol/L ( P<0.05) and the best effective period was the 3rd day ( P<0.05) in inducing hiPSCs to differentiate into cardiomyocytes. The optimal concentration of BMP-4 was 20.0 ng/mL ( P<0.05) and the best effective period was the 3rd day ( P<0.05). The relative expressions of ISL1, NKX2-5, MEF2C, MYL2, MYL7, and TNNT2 mRNAs, the positive expression ratio of cTnT detected by flow cytometry, and sarcomere structure detected by immunofluorescence staining of Wnt+BMP group were superior to those of Wnt group ( P<0.05). Conclusion: Joint regulation of Wnt and BMP signaling pathways can improve the differentiation efficiency of hiPSCs into cardiomyocytes.


Assuntos
Células-Tronco Pluripotentes Induzidas , Miócitos Cardíacos , Diferenciação Celular , Regulação da Expressão Gênica , Humanos , Transdução de Sinais
8.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi ; 34(10): 1332-1340, 2020 Oct 15.
Artigo em Chinês | MEDLINE | ID: mdl-33063501

RESUMO

Objective: To summarize the research progress of the regulatory role of microRNA (miRNA) in osteogenic differentiation of mesenchymal stem cells (MSCs) and its application as a therapeutic target and diagnostic tool in orthopedic diseases. Methods: The recent literature on the regulation of MSCs osteogenic differentiation by miRNAs was extensively reviewed, and its regulatory mechanism and its application as a therapeutic target and diagnostic tool in orthopedic diseases were reviewed. Results: miRNAs are small endogenous non-coding RNAs with a length of 20-22 nucleotides, which play an important role in the osteogenic differentiation of MSCs. Osteogenesis begins with the differentiation of MSCs into mature osteoblasts, and each stage of dynamic homeostasis of bone metabolism is associated with the regulation of different miRNAs. miRNAs are regulated from the post-transcriptional level by mRNAs cleavage, degradation, translational repression, or methylation. In addition, current studies suggest that miRNAs can be used as a new diagnostic tool and therapeutic target for orthopedic diseases. Conclusion: Further study on the regulation mechanism of miRNAs will provide more ideas for finding new therapeutic targets and diagnostic tools for orthopedic disease.


Assuntos
Células-Tronco Mesenquimais , MicroRNAs , Diferenciação Celular , MicroRNAs/genética , Osteoblastos , Osteogênese
9.
Nat Commun ; 11(1): 4989, 2020 10 05.
Artigo em Inglês | MEDLINE | ID: mdl-33020476

RESUMO

We postulate that exit from pluripotency involves intermediates that retain pluripotency while simultaneously exhibiting lineage-bias. Using a MIXL1 reporter, we explore mesoderm lineage-bias within the human pluripotent stem cell compartment. We identify a substate, which at the single cell level coexpresses pluripotent and mesodermal gene expression programmes. Functionally these cells initiate stem cell cultures and exhibit mesodermal bias in differentiation assays. By promoting mesodermal identity through manipulation of WNT signalling while preventing exit from pluripotency using lysophosphatidic acid, we 'trap' and maintain cells in a lineage-biased stem cell state through multiple passages. These cells correspond to a normal state on the differentiation trajectory, the plasticity of which is evidenced by their reacquisition of an unbiased state upon removal of differentiation cues. The use of 'cross-antagonistic' signalling to trap pluripotent stem cell intermediates with different lineage-bias may have general applicability in the efficient production of cells for regenerative medicine.


Assuntos
Reprogramação Celular , Mesoderma/metabolismo , Células-Tronco Pluripotentes/citologia , Animais , Diferenciação Celular/genética , Linhagem Celular , Linhagem da Célula , Plasticidade Celular/genética , Autorrenovação Celular , Meios de Cultura , Células-Tronco Embrionárias/citologia , Células-Tronco Embrionárias/metabolismo , Humanos , Camundongos , Células-Tronco Pluripotentes/metabolismo , Transdução de Sinais/genética
10.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 28(5): 1710-1717, 2020 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-33067979

RESUMO

AbstractObjective: To investigate the effect of ALAS2 downregulation on the expression of BNIP3L and erythroid differentiation in K562 cells. METHODS: The expression of ALAS2 was down-regulated by transfection of lentivirus, then quantitative real-time PCR was performed to detect the transfection efficiency. Flow cytometry analysis was applied to evaluate apoptosis of cells, erythroid differentiation, mitochondrial membrane potential and reactive oxygen species (ROS) level. Western blot was used to detect the BNIP3L expression, Co-immunoprecipitation was performed to analyze the relationship between ALAS2 and BNIP3L. RESULTS: Compared with sh-NC group, knockdown of ALAS2 induced downregulation of BNIP3L mRNA and protein expression(P<0.01) and erythroid related transcription factors GATA1, Nrf2 expression, as well as reduction of ROS level(P<0.05). Mitochondrial membrane potential of control (sh-NC) group was lower than that of shALAS2 group(P<0.05), but there was no significant change of cell apoptotic rate in two groups. CD71highCD235ahigh + CD71lowCD235ahigh cells of sh-NC and shALAS2 groups were 53.5%, 92.9% at 96 h after hemin induction, respectively. No direct action between ALAS2 and BNIP3L was observed. CONCLUSION: The intracellular heme level can affect the expression of BNIP3L which may be related with the regulation of ROS and transcription factors GATA1 and Nrf2. Higher BNIP3L facilitates cell differentiation but lower BNIP3L is favorable for cells survival.


Assuntos
Proteínas de Transporte , Mitofagia , 5-Aminolevulinato Sintetase/metabolismo , Apoptose , Diferenciação Celular , Humanos , Células K562 , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Proteínas Proto-Oncogênicas/genética , Proteínas Supressoras de Tumor
11.
Nat Commun ; 11(1): 4956, 2020 10 02.
Artigo em Inglês | MEDLINE | ID: mdl-33009383

RESUMO

Tet-enzyme-mediated 5-hydroxymethylation of cytosines in DNA plays a crucial role in mouse embryonic stem cells (ESCs). In RNA also, 5-hydroxymethylcytosine (5hmC) has recently been evidenced, but its physiological roles are still largely unknown. Here we show the contribution and function of this mark in mouse ESCs and differentiating embryoid bodies. Transcriptome-wide mapping in ESCs reveals hundreds of messenger RNAs marked by 5hmC at sites characterized by a defined unique consensus sequence and particular features. During differentiation a large number of transcripts, including many encoding key pluripotency-related factors (such as Eed and Jarid2), show decreased cytosine hydroxymethylation. Using Tet-knockout ESCs, we find Tet enzymes to be partly responsible for deposition of 5hmC in mRNA. A transcriptome-wide search further reveals mRNA targets to which Tet1 and Tet2 bind, at sites showing a topology similar to that of 5hmC sites. Tet-mediated RNA hydroxymethylation is found to reduce the stability of crucial pluripotency-promoting transcripts. We propose that RNA cytosine 5-hydroxymethylation by Tets is a mark of transcriptome flexibility, inextricably linked to the balance between pluripotency and lineage commitment.


Assuntos
5-Metilcitosina/análogos & derivados , Diferenciação Celular , Proteínas de Ligação a DNA/metabolismo , Células-Tronco Embrionárias Murinas/citologia , Células-Tronco Embrionárias Murinas/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , RNA/metabolismo , 5-Metilcitosina/metabolismo , Animais , Especificidade de Anticorpos/imunologia , Sequência de Bases , Corpos Embrioides/metabolismo , Camundongos , Modelos Biológicos , Células-Tronco Pluripotentes/metabolismo , Ligação Proteica , Estabilidade de RNA/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transcriptoma/genética
12.
Environ Health Prev Med ; 25(1): 59, 2020 Oct 08.
Artigo em Inglês | MEDLINE | ID: mdl-33032525

RESUMO

Asbestos exposure is known to cause malignant mesothelioma, which is associated with poor prognosis. We focused on and examined the effect of asbestos exposure on the differentiation and function of cytotoxic T lymphocytes (CTLs). CTLs have the ability to specifically attack tumor cells after being differentiated from naïve CD8+ T cells following antigen stimulation. Exposure to chrysotile B asbestos suppressed the differentiation of CTLs during the mixed lymphocyte reaction (MLR) and was associated with a decrease in proliferation of CD8+ T cells. Additionally, in an effort to investigate the mechanism associated with suppressed CTL differentiation upon exposure to asbestos, we focused on IL-2, a cytokine involved in T cell proliferation. Our findings indicated that insufficient levels of IL-2 are not the main cause for the suppressed induction of CTLs by asbestos exposure, although they suggest potential improvement in the suppressed CTL function. Furthermore, the functional properties of peripheral blood CD8+ lymphocytes from asbestos-exposed individuals with pleural plaque (PP) and patients with malignant mesothelioma (MM) were examined. MM patients showed lower perforin levels in CD8+ lymphocytes following stimulation compared with PP-positive individuals. The production capacity of IFN-γ in the MM group tended to be lower compared with healthy volunteers or PP-positive individuals. In an effort to determine whether chronic and direct asbestos exposure affected the function of CD8+ T cells, cultured human CD8+ T cells were employed as an in vitro model and subjected to long-term exposure to chrysotile (CH) asbestos. This resulted in decreased levels of intracellular perforin and secreted IFN-γ. Those findings underlie the possibility that impaired CD8+ lymphocyte function is caused by asbestos exposure, which fail to suppress the development of MM. Our studies therefore reveal novel effects of asbestos exposure on CTLs, which might contribute towards the development and implementation of an effective strategy for the prevention and cure of malignant mesothelioma.


Assuntos
Asbestos/toxicidade , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Neoplasias Pulmonares/imunologia , Mesotelioma/imunologia , Linfócitos T Citotóxicos/efeitos dos fármacos , Asbestos Serpentinas/toxicidade , Humanos , Linfócitos T Citotóxicos/imunologia
13.
Nat Commun ; 11(1): 5063, 2020 10 08.
Artigo em Inglês | MEDLINE | ID: mdl-33033242

RESUMO

Genome-wide chromatin state underlies gene expression potential and cellular function. Epigenetic features and nucleosome positioning contribute to the accessibility of DNA, but widespread regulators of chromatin state are largely unknown. Our study investigates how coordination of ANP32E and H2A.Z contributes to genome-wide chromatin state in mouse fibroblasts. We define H2A.Z as a universal chromatin accessibility factor, and demonstrate that ANP32E antagonizes H2A.Z accumulation to restrict chromatin accessibility genome-wide. In the absence of ANP32E, H2A.Z accumulates at promoters in a hierarchical manner. H2A.Z initially localizes downstream of the transcription start site, and if H2A.Z is already present downstream, additional H2A.Z accumulates upstream. This hierarchical H2A.Z accumulation coincides with improved nucleosome positioning, heightened transcription factor binding, and increased expression of neighboring genes. Thus, ANP32E dramatically influences genome-wide chromatin accessibility through subtle refinement of H2A.Z patterns, providing a means to reprogram chromatin state and to hone gene expression levels.


Assuntos
Cromatina/metabolismo , Genoma , Chaperonas Moleculares/metabolismo , Animais , Diferenciação Celular/genética , DNA Helicases/metabolismo , Embrião de Mamíferos/metabolismo , Fibroblastos/citologia , Fibroblastos/metabolismo , Regulação da Expressão Gênica , Histonas/metabolismo , Camundongos , Proteínas Nucleares/metabolismo , Nucleossomos/metabolismo , Regiões Promotoras Genéticas , Ligação Proteica , Fatores de Transcrição/metabolismo
14.
Annu Int Conf IEEE Eng Med Biol Soc ; 2020: 2239-2244, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-33018453

RESUMO

Directing the fate of human mesenchymal stem/stromal cells (hMSCs) toward bone formation using mechanical strain is a promising approach in regenerative medicine related to bone diseases. Numerous studies have evaluated the effects of vibration or cyclic tensile strain on MSCs towards developing a mechanically-based method for stimulating differentiation. Here, we study the differentiation of hMSCs cultured on elastic polydimethylsiloxane (PDMS) membrane, which is magnetically actuated to induce periodically varying strain. The strain distribution across the membrane was calculated by finite-element modeling and demonstrates three main areas of different strain amplitudes. The strain effect on the hMSCs was evaluated by measuring the mineralization of differentiated hMSCs using Alizarin S red stain. The results indicate a strain-dependent differentiation of hMSCs, where the highest region of strain on the membrane resulted in the most accelerated differentiation. Osteogenic differentiation was achieved as early as two weeks, which is significantly sooner than control hMSCs treated with osteogenic media alone.


Assuntos
Células-Tronco Mesenquimais , Diferenciação Celular , Humanos , Osteogênese
15.
Folia Biol (Praha) ; 66(3): 111-115, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33069190

RESUMO

Progenitor cells of the human erythroid and granulocytic cell lineages are characterized by the presence of several nucleoli. One of these nucleoli is larger and possesses more fibrillar centres than others. Such nucleolus is apparently dominant in respect of both size and main nucleolar function such as nucleolar-ribosomal RNA transcription. Such nucleolus is also visible in specimens using conventional visualization procedures, in contrast to smaller nucleoli. In the terminal differentiation nucleated stages of the erythroid and granulocytic development, dominant nucleoli apparently disappeared, since these cells mostly contained very small nucleoli of a similar size with one fibrillar centre. Thus, the easily visible dominant nucleoli appear to be useful markers of the progenitor cell state, such as proliferation, and differentiation potential.


Assuntos
Nucléolo Celular/fisiologia , Células Precursoras Eritroides/ultraestrutura , Células Precursoras de Granulócitos/ultraestrutura , Diferenciação Celular , Divisão Celular , Linhagem da Célula , Nucléolo Celular/ultraestrutura , Núcleo Celular/ultraestrutura , Granulócitos/ultraestrutura , Humanos , RNA Ribossômico/metabolismo
16.
Shanghai Kou Qiang Yi Xue ; 29(4): 343-349, 2020 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-33089279

RESUMO

PURPOSE: To investigate the biological characteristics of human periodontal stem cells (hPDLSCs) modified with platelet derived growth factor BB(PDGFBB) gene, and to explore its influence on proliferation, migration and osteogenic induction of hPDLSCs. METHODS: hPDLSCs were isolated and amplified, and immunofluorescence staining was performed to identify cell surface markers and osteogenic differentiation ability. hPDLSCs were transfected with PDGFBB gene by lentivirus vector, and the effects on cell proliferation and migration were detected by CCK-8 and scratch test after transfection. Real-time PCR was performed to analyze the mRNA expression levels of osteogenic and angiogenic genes in hPDLSCs cells transfected with PDGFBB gene. Statistical analysis was performed using SPSS 22.0 software package. RESULTS: hPDLSCs were successfully obtained by tissue mass culture and finite dilution method. Compared with the blank virus group and non-transfected group, the proliferation and migration ability of the cells in the transfection group were significantly increased, and the mRNA expression levels of OPN, COL-1 and VEGF were significantly up-regulated(P<0.05). CONCLUSIONS: Lentiviral vector can transfer PDGFBB gene into hPDLSCs in vitro and obtain continuous and stable expression. PDGFBB can promote proliferation and migration of hPDLSCs cells and up-regulate expression of osteogenic and angiogenic genes.


Assuntos
Becaplermina , Ligamento Periodontal , Diferenciação Celular , Humanos , Osteogênese/genética , Células-Tronco
17.
J Toxicol Sci ; 45(9): 569-579, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32879256

RESUMO

Indoxyl, a derivative of indole originating from tryptophan, may undergo phase-II sulfate-conjugation pathway, thereby forming indoxyl sulfate (IS) in vivo. We previously reported that IS, a well-known uremic toxin, can increase the intracellular oxidation level and decrease the phagocytic activity in a differentiated HL-60 human macrophage cell model. Using the same cell model, the current study aimed to investigate whether indole and indoxyl (the metabolic precursors of indoxyl and IS, respectively) may cause macrophage immune dysfunction. Results obtained indicated that intracellular oxidation level and cytotoxicity markedly increased upon treatment with indole and indoxyl, in comparison with IS. Incubation of the cells with indole and indoxyl also resulted in attenuated phagocytic activity. Human serum albumin (HSA)-binding assay confirmed that tryptophan and IS, but not indole and indoxyl, could selectively bind to the site II in HSA. Collectively, the results indicated that indole and indoxyl may strongly down-regulate the phagocytic immune function of macrophages, whereas IS, formed upon sulfate conjugation of indoxyl, may exhibit enhanced HSA-binding capability, thereby reducing the adverse effects of indoxyl.


Assuntos
Indóis/efeitos adversos , Macrófagos/imunologia , Macrófagos/metabolismo , Oxirredução/efeitos dos fármacos , Fagocitose/efeitos dos fármacos , Fagocitose/imunologia , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Células HL-60 , Humanos , Indicã/metabolismo , Macrófagos/efeitos dos fármacos , Ligação Proteica , Albumina Sérica/metabolismo , Triptofano/metabolismo
18.
Nat Commun ; 11(1): 4786, 2020 09 22.
Artigo em Inglês | MEDLINE | ID: mdl-32963227

RESUMO

Evidence points to an indispensable function of macrophages in tissue regeneration, yet the underlying molecular mechanisms remain elusive. Here we demonstrate a protective function for the IL-33-ST2 axis in bronchial epithelial repair, and implicate ST2 in myeloid cell differentiation. ST2 deficiency in mice leads to reduced lung myeloid cell infiltration, abnormal alternatively activated macrophage (AAM) function, and impaired epithelial repair post naphthalene-induced injury. Reconstitution of wild type (WT) AAMs to ST2-deficient mice completely restores bronchial re-epithelialization. Central to this mechanism is the direct effect of IL-33-ST2 signaling on monocyte/macrophage differentiation, self-renewal and repairing ability, as evidenced by the downregulation of key pathways regulating myeloid cell cycle, maturation and regenerative function of the epithelial niche in ST2-/- mice. Thus, the IL-33-ST2 axis controls epithelial niche regeneration by activating a large multi-cellular circuit, including monocyte differentiation into competent repairing AAMs, as well as group-2 innate lymphoid cell (ILC2)-mediated AAM activation.


Assuntos
Bronquíolos/metabolismo , Diferenciação Celular/efeitos dos fármacos , Células Epiteliais/metabolismo , Proteína 1 Semelhante a Receptor de Interleucina-1/metabolismo , Interleucina-33/metabolismo , Interleucina-33/farmacologia , Animais , Bronquíolos/lesões , Bronquíolos/patologia , Citocinas/metabolismo , Modelos Animais de Doenças , Células Epiteliais/patologia , Feminino , Proteína 1 Semelhante a Receptor de Interleucina-1/genética , Pulmão/patologia , Ativação Linfocitária , Linfócitos/metabolismo , Macrófagos/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Transdução de Sinais
19.
Nat Commun ; 11(1): 4788, 2020 09 22.
Artigo em Inglês | MEDLINE | ID: mdl-32963234

RESUMO

Regenerative proliferation capacity and poor differentiation are histological features usually linked to poor prognosis in head and neck squamous cell carcinoma (hnSCC). However, the pathways that regulate them remain ill-characterized. Here, we show that those traits can be triggered by the RHO GTPase activator VAV2 in keratinocytes present in the skin and oral mucosa. VAV2 is also required to maintain those traits in hnSCC patient-derived cells. This function, which is both catalysis- and RHO GTPase-dependent, is mediated by c-Myc- and YAP/TAZ-dependent transcriptomal programs associated with regenerative proliferation and cell undifferentiation, respectively. High levels of VAV2 transcripts and VAV2-regulated gene signatures are both associated with poor hnSCC patient prognosis. These results unveil a druggable pathway linked to the malignancy of specific SCC subtypes.


Assuntos
Proliferação de Células , Neoplasias de Cabeça e Pescoço/metabolismo , Proteínas Proto-Oncogênicas c-vav/genética , Proteínas Proto-Oncogênicas c-vav/metabolismo , Carcinoma de Células Escamosas de Cabeça e Pescoço/metabolismo , Animais , Diferenciação Celular , Modelos Animais de Doenças , Epiderme/metabolismo , Epiderme/patologia , GTP Fosfo-Hidrolases , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Neoplasias de Cabeça e Pescoço/genética , Neoplasias de Cabeça e Pescoço/patologia , Humanos , Hiperplasia/patologia , Queratinócitos/patologia , Camundongos , Camundongos Knockout , Membrana Mucosa/metabolismo , Prognóstico , RNA Mensageiro/metabolismo , Transdução de Sinais , Carcinoma de Células Escamosas de Cabeça e Pescoço/genética , Carcinoma de Células Escamosas de Cabeça e Pescoço/patologia , Transcriptoma
20.
Nat Commun ; 11(1): 4816, 2020 09 23.
Artigo em Inglês | MEDLINE | ID: mdl-32968047

RESUMO

Understanding cell types and mechanisms of dental growth is essential for reconstruction and engineering of teeth. Therefore, we investigated cellular composition of growing and non-growing mouse and human teeth. As a result, we report an unappreciated cellular complexity of the continuously-growing mouse incisor, which suggests a coherent model of cell dynamics enabling unarrested growth. This model relies on spatially-restricted stem, progenitor and differentiated populations in the epithelial and mesenchymal compartments underlying the coordinated expansion of two major branches of pulpal cells and diverse epithelial subtypes. Further comparisons of human and mouse teeth yield both parallelisms and differences in tissue heterogeneity and highlight the specifics behind growing and non-growing modes. Despite being similar at a coarse level, mouse and human teeth reveal molecular differences and species-specific cell subtypes suggesting possible evolutionary divergence. Overall, here we provide an atlas of human and mouse teeth with a focus on growth and differentiation.


Assuntos
Diferenciação Celular , Células-Tronco/citologia , Dente/citologia , Dente/crescimento & desenvolvimento , Adolescente , Adulto , Animais , Diferenciação Celular/genética , Células Epiteliais , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Heterogeneidade Genética , Humanos , Incisivo/citologia , Incisivo/crescimento & desenvolvimento , Masculino , Mesoderma/citologia , Mesoderma/crescimento & desenvolvimento , Mesoderma/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Modelos Animais , Dente Molar/citologia , Dente Molar/crescimento & desenvolvimento , Odontoblastos , Adulto Jovem
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