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The importance of incorporating an agile approach into creating sustainable products has been widely discussed. This approach can enhance innovation integration, improve adaptability to changing development circumstances, and increase the efficiency and quality of the product development process. While many agile methods have originated in the software development context and have been formulated based on successful software projects, they often fail due to incorrect procedures and a lack of acceptance, preventing deep integration into the process. Additionally, decision-making for market evaluation is often hindered by unclear and subjective information. Therefore, this study introduces an extended TOPSIS (Technique for Order Performance by Similarity to Ideal Solution) method for sustainable product development. This method leverages the benefits of cloud model theory to address randomness and uncertainty (intrapersonal uncertainty) and the advantages of rough set theory to flexibly handle market demand uncertainty without requiring extra information. The study proposes an integrated weighting method that considers both subjective and objective weights to determine comprehensive criteria weights. It also presents a new framework, named Sustainable Agility of Product Development (SAPD), which aims to evaluate criteria for assessing sustainable product development. To validate the effectiveness of this proposed method, a case study is conducted on small and medium enterprises in China. The obtained results show that the company needs to conduct product structure research and development to realize new product functions.
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Desenvolvimento Sustentável , China , Incerteza , Humanos , Computação em Nuvem , Software , Modelos TeóricosRESUMO
To solve the problems of poor top coal drawing, lagging collapse, and difficulty in parallel operation of mining and drawing in extra-thick coal seams, considering the 8222 working face of the Tashan Mine as the engineering background, we first investigate the basic problems of fully mechanized top coal drawing mining in extra-thick coal seams using the single-round group drawing method (GDM). We then analyze the drawing law of top coal with different thicknesses under the single-round GDM from the aspects of top coal recovery (TCR) and drawing efficiency, coal loss mechanism, and the relation between TCR and gangue content (GC), providing a basis for determining the process parameters of GDM. Results indicate that as the top coal thickness increases, the number of drawing openings considerably influences drawing efficiency and top coal loss. And there is a notable thickness effect of the number of drawing openings on the top coal loss. There is a quantitative relationship among TCR, cumulative GC (CGC) and instantaneous GC (IGC), and CGC and TCR can be predicted based on the IGC. Consequently, theoretical analysis and numerical simulation results indicate that the optimal IGC threshold at the coal drawing openings between 31.2 and 40%. Through optimizing the coal drawing method and strictly controlling the IGC at the coal drawing openings on-site, the measured working face TCR increased from 75.25 to 90.12%, and CGC was controlled at approximately 9%. Meanwhile, the average coordination efficiency of mining and drawing time reaches 68.2%, effectively ensuring the construction of a coal mine with an annual output of 15 million tons.
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Lower extremity deep venous thrombosis (LEDVT) affects patient's quality of life for a long time, and even causes pulmonary embolism, which threatens human health. Current anticoagulant drugs in clinical treatment are hampered by the risk of bleeding due to poor targeting and low drug penetration. Here, we used platelet (PLT)-like biological targeting to enhance the delivery and accumulation of nanomedicines in thrombus and reduce the risk of bleeding. Meanwhile, the parallel strategy of "thrombus thermal ablation and anticoagulation" was applied to increase the permeability of drugs in thrombus and achieve the optimal antithrombotic effect. Polypyrrole (PPy) and rivaroxban (Riv, an anticoagulant drug) were co-assembled into platelet membrane-coated nanoparticles (NPs), PLT-PPy/Riv NPs, which actively targeted the thrombotic lesion at multiple targets in the platelet membrane and were thermally and drug-specific thrombolysed by 808 nm laser irradiation. The combination therapy resulted in up to 90% thrombolysis in a femoral vein thrombosis model compared to single phototherapy or drug therapy. The results showed that the nanoformulation provided a new direction for remote precise and controlled sustained thrombolysis, which was in line with the trend of nanomedicine towards clinical translation.
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Nanopartículas , Trombose , Trombose Venosa , Humanos , Polímeros/uso terapêutico , Fibrinolíticos/uso terapêutico , Pirróis/uso terapêutico , Preparações Farmacêuticas , Biomimética , Qualidade de Vida , Trombose Venosa/tratamento farmacológico , Trombose/tratamento farmacológico , Nanopartículas/uso terapêuticoRESUMO
[This corrects the article DOI: 10.1021/acsomega.3c01289.].
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Periodontitis is a progressive inflammatory skeletal disease characterized by periodontal tissue destruction, alveolar bone resorption, and tooth loss. Chronic inflammatory response and excessive osteoclastogenesis play essential roles in periodontitis progression. Unfortunately, the pathogenesis that contributes to periodontitis remains unclear. As a specific inhibitor of the mTOR (mammalian/mechanistic target of rapamycin) signaling pathway and the most common autophagy activator, rapamycin plays a vital role in regulating various cellular processes. The present study investigated the effects of rapamycin on osteoclast (OC) formation in vitro and its effects on the rat periodontitis model. The results showed that rapamycin inhibited OC formation in a dose-dependent manner by up-regulating the Nrf2/GCLC signaling pathway, thus suppressing the intracellular redox status, as measured by 2',7'-dichlorofluorescein diacetate and MitoSOX. In addition, rather than simply increasing the autophagosome formation, rapamycin increased the autophagy flux during OC formation. Importantly, the anti-oxidative effect of rapamycin was regulated by an increase in autophagy flux, which could be attenuated by blocking autophagy with bafilomycin A1. In line with the in vitro results, rapamycin treatment attenuated alveolar bone resorption in rats with lipopolysaccharide-induced periodontitis in a dose-dependent manner, as assessed by micro-computed tomography, hematoxylin-eosin staining, and tartrate-resistant acid phosphatase staining. Besides, high-dose rapamycin treatment could reduce the serum levels of proinflammatory factors and oxidative stress in periodontitis rats. In conclusion, this study expanded our understanding of rapamycin's role in OC formation and protection from inflammatory bone diseases.
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OBJECTIVE: This study aims to investigate a novel pathogenic variant in a Chinese family of non-syndromic tooth agenesis (NSTA) and study the impact of the variant on related protein and pathway. DESIGN: One NSTA family was collected. Whole exome sequencing and Sanger sequencing were performed on the proband with NSTA and his 5 family members. The pathogenic influence of the mutant is evaluated by bioinformatics analyses including evolutionary conservation analysis and secondary structure prediction. Molecular dynamics (MD) simulations and binding free energy calculations were then performed to explore changes in the tertiary structure and binding ability of the protein. RESULTS: We found a novel missense ectodysplasin A receptor (EDAR) variant (c .1292 T > G; p.Ile431Arg) in all affected family members. The results of bioinformatics analyses revealed that the EDAR had harmful changes after mutation. MD simulations and the binding free energy calculations results showed that the mutant EDAR protein and EDAR/ectodysplasin-A receptor-associated adapter (EDARADD) complex displayed tertiary structural change, and EDAR possessed a lower affinity to EDARADD after mutation. CONCLUSIONS: We found a novel EDAR variant (c.1292 T > G; p.Ile431Arg) in one NSTA family, which affects the binding of EDAR and EDARADD.
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Anodontia , Simulação de Dinâmica Molecular , Humanos , Receptores da Ectodisplasina/genética , Anodontia/genética , Mutação , Proteínas Wnt/genética , Receptor Edar/genética , Ectodisplasinas/genéticaRESUMO
Triple-negative breast cancer (TNBC) is a group of fatal malignancies characterized by high metastatic capacity, the underlying mechanisms of which remain largely elusive. We have found here that insulin-like growth factor 2 mRNA binding protein 3 (IGF2BP3) is highly expressed in TNBC and correlates clinically with distant metastasis-free survival of TNBC patients. IGF2BP3 promotes the migration and invasion capabilities of TNBC cells dependent upon cellular RNA N6-methyladenosine (m6A) modification. Mechanistically, IGF2BP3 binds to and destabilizes m6A-methylated mRNA of the extracellular matrix glycoprotein, SLIT2, impairs its downstream signaling via the cognate receptor ROBO1, and consequently triggers the activation of canonical PI3K/AKT and MEK/ERK pathways. The IGF2BP3/SLIT2 axis is critically involved in the regulation of TNBC metastasis in vivo. These findings shed light into the regulatory network of distant metastasis of breast cancer and provide rationale for targeting the m6A machinery in the treatment of TNBC.
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Proteínas de Ligação a RNA , Somatomedinas , Neoplasias de Mama Triplo Negativas , Humanos , Glicoproteínas , Quinases de Proteína Quinase Ativadas por Mitógeno , Proteínas do Tecido Nervoso/genética , Fosfatidilinositol 3-Quinases , Proteínas Proto-Oncogênicas c-akt , Receptores Imunológicos/genética , RNA , RNA Mensageiro/genética , Neoplasias de Mama Triplo Negativas/metabolismo , Proteínas de Ligação a RNA/genética , Peptídeos e Proteínas de Sinalização Intercelular/genéticaRESUMO
As an emerging novel drug delivery system, microneedles (MNs) have a wide range of applications in the medical field. They can overcome the physiological barriers of the skin, penetrate the outermost skin of the human body, and form hundreds of reversible microchannels to enhance the penetration of drugs and deliver drugs to the diseased sites. So they have great applications in the diagnosis and treatment of melanoma. Melanoma is a kind of malignant tumor, the survival rate of patients with metastases is extremely low. The traditional methods of surgery and drug treatment for melanoma are often accompanied by large adverse reactions in the whole body, and the drug concentration is low. The use of MNs for transdermal administration can increase the drug concentration, reduce adverse reactions in the treatment process, and have good therapeutic effect on melanoma. This paper introduced various types of MNs and their preparation methods, summarized the diagnosis and various treatment options for melanoma with MNs, focused on the treatment of melanoma with dissolved MNs, and made prospect of MNs-mediated transdermal drug delivery in the treatment of melanoma.
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Melanoma is an aggressive malignancy deriving from melanocytes, which is characterized by high tendency of metastases and mortality rate. Current therapies for melanoma, like chemotherapy, immunotherapy and targeted therapy, have the problem of systemic exposure of drugs, which will lead to many side effects and premature degradation of drugs. The resulting low drug accumulation at the lesion limits the therapeutic effect on melanoma and makes the cure rate low. As an emerging drug delivery system, microneedles (MNs) can efficiently deliver drugs through the skin, increase the drug distribution in deeper tumor sites and minimize the leakage of therapeutic drugs into adjacent tissues, thus improving the therapeutic effect. In addition, compared with traditional drug delivery methods, MN-based drug delivery system has the advantages of simplicity, safety and little pain. So MNs can be developed for the treatment of melanoma, which can relieve the pain of patients and improve the survival rate. This review aims to introduce an update on the progress of MNs as an innovative strategy for melanoma, especially when MNs combining with different therapies against melanoma, such as chemotherapy, targeted therapy, immunotherapy, photothermal therapy (PTT), photodynamic therapy (PDT) and synergic therapy.
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Melanoma , Neoplasias Cutâneas , Administração Cutânea , Sistemas de Liberação de Medicamentos/métodos , Humanos , Melanoma/tratamento farmacológico , Dor , Neoplasias Cutâneas/tratamento farmacológico , Neoplasias Cutâneas/patologiaRESUMO
Integrated treatment using imaging technology to monitor biological processes for the precise treatment and diagnosis of diseases to improve treatment outcomes is becoming a hot topic. Accordingly, perylene diimide (PDI) has excellent photothermal conversion and photostability, which can be used as a good material for disease treatment and diagnosis. Herein, we review the latest research progress on the real-time diagnosis of related diseases based on perylene diimide probes in the aspects of bioimaging, detection of biomarkers and determination of the pH in living cells. Furthermore, perylene diimide-based multifunctional nano-delivery systems are particularly emphasized, showing great therapeutic potential in the field of image-guided combination therapy in tumor therapy. Finally, the great opportunities and challenges still faced by perylene diimide before entering the clinic are comprehensively analyzed.
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Antineoplásicos/uso terapêutico , Imidas/uso terapêutico , Neoplasias/tratamento farmacológico , Perileno/análogos & derivados , Técnicas Fotoacústicas , Antineoplásicos/química , Humanos , Imidas/química , Neoplasias/diagnóstico , Tamanho da Partícula , Perileno/química , Perileno/uso terapêutico , Propriedades de SuperfícieRESUMO
BACKGROUND: Although ongoing development of therapeutic strategies contributes to the improvements in clinical management, clear cell renal cell carcinoma (ccRCC) deaths originate mainly from radiochemoresistant and metastatic disease. Transcription factor SALL4 has been implicated in tumorigenesis and metastasis of multiple cancers. However, it is not known whether SALL4 is involved in the pathogenesis of ccRCC. METHODS: Analyses of clinical specimen and publicly available datasets were performed to determine the expression level and clinical significance of SALL4 in ccRCC. The influence of SALL4 expression on ccRCC tumor growth, metastasis and vascularity was evaluated through a series of in vitro and in vivo experiments. Western blotting, immunofluorescence staining and integrative database analysis were carried out to investigate the underlying mechanism for SALL4-mediated oncogenic activities in ccRCC. RESULTS: SALL4 expression was increased in ccRCC and positively correlated with tumor progression and poor prognosis. SALL4 could promote ccRCC cell proliferation, colony formation, cell cycle progression, migration, invasion and tumorigenicity and inhibit cell senescence. Further investigation revealed a widespread association of SALL4 with individual gene transcription and the involvement of SALL4 in endothelium development and vasculogenesis. In the context of ccRCC, SALL4 promoted tumor vascularization by recruiting endothelial cells. In addition, we found that SALL4 could exert its tumor-promoting effect via modulating Akt/GSK-3ß axis and VEGFA expression. VHL mutation and DNA hypomethylation may be involved in the upregulation of SALL4 in ccRCC. CONCLUSIONS: Overall, our results provide evidence that upregulated SALL4 can function as a crucial regulator of tumor pathogenesis and progression in ccRCC, thus offering potential therapeutic strategies for future treatment.
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Carcinoma de Células Renais/patologia , Glicogênio Sintase Quinase 3 beta/metabolismo , Neoplasias Renais/patologia , Mutação , Proteínas Proto-Oncogênicas c-akt/metabolismo , Fatores de Transcrição/metabolismo , Proteína Supressora de Tumor Von Hippel-Lindau/genética , Animais , Apoptose , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Carcinoma de Células Renais/irrigação sanguínea , Carcinoma de Células Renais/genética , Carcinoma de Células Renais/metabolismo , Proliferação de Células , Progressão da Doença , Feminino , Regulação Neoplásica da Expressão Gênica , Glicogênio Sintase Quinase 3 beta/genética , Humanos , Neoplasias Renais/irrigação sanguínea , Neoplasias Renais/genética , Neoplasias Renais/metabolismo , Masculino , Camundongos , Camundongos Nus , Pessoa de Meia-Idade , Invasividade Neoplásica , Neovascularização Patológica , Prognóstico , Proteínas Proto-Oncogênicas c-akt/genética , Transdução de Sinais , Taxa de Sobrevida , Fatores de Transcrição/genética , Células Tumorais Cultivadas , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
BACKGROUND Spalt-like protein 4 (SALL4) is a nuclear transcription factor central to early embryonic development, especially for regulating pluripotency of embryonic stem cells (ESCs) and sustaining ESCs self-renewal. Aberrant re-expression of SALL4 in adult tissues is involved in tumorigenesis and cancer progression. However, the role of SALL4 in angiogenesis remains elusive. Here, we determined the potential action of SALL4 on proliferation, migration, and tube formation of endothelial cells. MATERIAL AND METHODS HUVECs were infected with lentiviral particles expressing shRNA against SALL4. QRT-PCR and immunoblotting analysis were carried out to evaluate knockdown efficiency at mRNA and protein levels. Cell proliferation was measured by CCK-8 assay and flow cytometry was conducted to analyze cell cycle distribution. Wound-healing and Transwell migration assays were performed to evaluate cell motility. In addition, we determined the role of SALL4 on angiogenesis by tube formation assay, and Western blot analysis was used to assess the effect of SALL4 downregulation on VEGFA expression. RESULTS We found that SALL4 downregulation resulted in decreased proliferation. Cell cycle analysis revealed that SALL4 knockdown impeded cell cycle progression and induced cell cycle arrest at G1 phase. We also found that silencing of SALL4 decreased the capacity of wound healing and cell migration in HUVECs. Furthermore, tube formation assay showed that loss of SALL4 inhibited HUVECs angiogenesis. We also observed that SALL4 knockdown reduced the level of VEGFA in HUVECs. CONCLUSIONS In conclusion, these results support that by promoting proliferation, cell cycle progression, migration, and tube formation, SALL4 is involved in the process of angiogenesis through modulating VEGFA expression.
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Movimento Celular/fisiologia , Proliferação de Células/fisiologia , Células Endoteliais/fisiologia , Neovascularização Fisiológica/fisiologia , Fatores de Transcrição/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Proteínas Angiogênicas/metabolismo , Fase G1 , Regulação da Expressão Gênica , Células Endoteliais da Veia Umbilical Humana , Humanos , Transdução de SinaisRESUMO
Epigenetic alterations that lead to dysregulated gene expression in the progression of castration-resistant prostate cancer (CRPC) remain elusive. Here, we investigated the role of histone deubiquitinase MYSM1 in the pathogenesis of prostate cancer (PCa). Tissues and public datasets of PCa were evaluated for MYSM1 levels. We explored the effects of MYSM1 on cell proliferation, senescence and viability both in vitro and in vivo. Integrative database analyses and co-immunoprecipitation assays were performed to elucidate genomic association of MYSM1 and MYSM1-involved biological interaction network in PCa. We observed that MYSM1 were downregulated in CRPC compared to localized prostate tumors. Knockdown of MYSM1 promoted cell proliferation and suppressed senescence of CRPC cells under condition of androgen ablation. MYSM1 downregulation enhanced the tumorigenic ability in nude mice. Integrative bioinformatic analyses of the significantly associated genes with MYSM1 revealed MYSM1-correlated pathways, providing substantial clues as to the role of MYSM1 in PCa. MYSM1 was able to bind to androgen receptor instead of increasing its expression and knockdown of MYSM1 resulted in activation of Akt/c-Raf/GSK-3ß signaling. Together, our findings indicate that MYSM1 is pivotal in CRPC pathogenesis and may be established as a potential target for future treatment.
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Regulação Neoplásica da Expressão Gênica/fisiologia , Neoplasias de Próstata Resistentes à Castração/patologia , Receptores Androgênicos/metabolismo , Transdução de Sinais/fisiologia , Transativadores/metabolismo , Proteases Específicas de Ubiquitina/metabolismo , Animais , Glicogênio Sintase Quinase 3 beta/metabolismo , Humanos , Masculino , Camundongos , Camundongos Nus , Proteína Oncogênica v-akt/metabolismo , Neoplasias de Próstata Resistentes à Castração/metabolismo , Proteínas Proto-Oncogênicas c-raf/metabolismoRESUMO
Prostate cancer (PCa) is a common malignancy in male urinary system. Cell division cycle-associated protein 1 (CDCA1) is expressed highly in many cancer cells. Yet, whether CDCA1 play an important role in PCa progression is uncertain. pH low insertion peptide (pHLIP), a PH-induced transmembrane structure, can pass through the cell membrane into intracellular in an acidic environment. In this study, we try to confirm the expression status of CDCA1 in the PCa patients' tissues and PCa cell line. In addition, to make the CDCA1-siRNA efficiently targeting the PCa cells, pHLIP and CDCA1-siRNA were combined with disulfide bond to become effector molecules. By the characteristics of the pHLIP allosteric occurring in cancer tissue acidic microenvironment, CDCA1-siRNA may be transported specificity into prostatic cancer cells and released in the cytoplasm. The interference effect of the effector molecules on the CDCA1 was detected in vitro and in vivo. The results showed that CDCA1 was highly expressed in PCa cell line and human PCa clinical samples. Knock down CDCA1 significantly inhibit the growth and promote the apoptosis of prostatic cancer cells. In the intracellular translocation experiment, CDCA1-siRNA could be delivered into cytoplasma at pH 6.2, but not at pH 7.4. In the in vivo test, the tumor size was reduced obviously in the NOD/SCID mice treated with pHLIP-CDCA1-siRNA compared to the CDCA1-siRNA and the bioluminescent signal of Cy5-pHLIP-CDCA1-siRNA was focused detected in the tumor site. Our findings indicated that CDCA1 might be a very key molecule regulating survival and proliferation of PCa. pHLIP-CDCA1-siRNA might be a promising targeting therapy for PCa.
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Proteínas de Ciclo Celular/genética , Neoplasias da Próstata/terapia , RNA Interferente Pequeno/metabolismo , Microambiente Tumoral , Animais , Células Cultivadas , Terapia Genética , Humanos , Concentração de Íons de Hidrogênio , Masculino , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Neoplasias Experimentais/metabolismo , Neoplasias Experimentais/patologia , Neoplasias Experimentais/terapia , Peptídeos , Neoplasias da Próstata/genética , Neoplasias da Próstata/patologia , RNA Interferente Pequeno/genéticaRESUMO
Targeted therapy is needed for hepatitis B virus (HBV)-mediated hepatocellular carcinoma (HCC) which shows overexpression of HBV surface antigen (HBsAg). We previously developed scFv15, a human single-chain antibody against HBsAg. Here we tested the strategic feasibility of scFv15-mediated delivery of apoptotic effectors for HBsAg-targeted HCC therapy and application of HA2 motif of influenza hemagglutinin to enhance endosome escape and antitumor effect. A class of HBsAg-targeted immunoproapoptotic molecule was generated by sequentially fusing scFv15, the furin-cleavable motif from diphtheria toxin (Fdt), HA2 and a truncated apoptotic protein Bid (tBid). The resulting scFv15-Fdt-HA2-tBid was prokaryotically expressed and functionally characterized for HBsAg-binding capacity, endosome escape activity and antitumor effect as compared with scFv15-Fdt-tBid. Both scFv15-Fdt-HA2-tBid and scFv15-Fdt-tBid retained affinity and specificity for HBsAg, and bound and selectively killed HBsAg-positive HCC cells via apoptosis. Notably, the IC50 of scFv15-Fdt-HA2-tBid in HBsAg-positive PLC/PRF/5 cells was 10 times lower than that of scFv15-Fdt-tBid. In vivo imaging of antitumor activity demonstrated 95% growth inhibition of orthotopic HCC by scFv15-Fdt-HA2-tBid compared with 75% suppression by scFv15-Fdt-tBid. This study represents an extended application of the immunoproapoptotic strategy in the treatment of HBsAg-positive HCC and shows significant potential of HA2 as a functional enhancer for endosome-encapsulated antibody-conjugates.
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Proteína Agonista de Morte Celular de Domínio Interatuante com BH3/metabolismo , Carcinoma Hepatocelular/tratamento farmacológico , Antígenos de Superfície da Hepatite B/imunologia , Vírus da Hepatite B/fisiologia , Neoplasias Hepáticas/tratamento farmacológico , Proteínas Recombinantes de Fusão/uso terapêutico , Anticorpos de Cadeia Única/uso terapêutico , Idoso , Sequência de Aminoácidos , Animais , Apoptose/efeitos dos fármacos , Carcinoma Hepatocelular/patologia , Carcinoma Hepatocelular/virologia , Linhagem Celular Tumoral , Endossomos/efeitos dos fármacos , Endossomos/metabolismo , Feminino , Fluoresceína/metabolismo , Vírus da Hepatite B/efeitos dos fármacos , Humanos , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/virologia , Masculino , Camundongos , Pessoa de Meia-Idade , Dados de Sequência Molecular , Ligação Proteica/efeitos dos fármacos , Transporte Proteico/efeitos dos fármacos , Proteínas Recombinantes de Fusão/farmacologia , Anticorpos de Cadeia Única/farmacologiaRESUMO
Although tamoxifen (TAM), a selective estrogen receptor modulator, has been widely used in the treatment of hormone-responsive breast cancer, its estrogen-like effect increases the risk of endometrial cancer. However, the molecular mechanisms of TAM-induced endometrial carcinoma still remain unclear. In this report, we explored the role of microRNAs (miRNAs) in TAM-induced epithelial-mesenchymal transition (EMT) in ECC-1 and Ishikawa endometrial cancer cell lines and found miR-200 is involved in this process via the regulation of c-Myc. When treated with TAM, ECC-1 and Ishikawa cells were characterized by higher invasiveness and motility and underwent EMT. miR-200, a miRNA family with tumor suppressive functions in a wide range of cancers, was found reduced in response to TAM treatment. Consistent with zinc finger E-box binding homeobox 2, which was confirmed as a direct target of miR-200b in endometrial cancer cell lines, some other key factors of EMT such as Snail and N-cadherin increased, whereas E-cadherin decreased in the TAM-treated cells, contributing to TAM-induced EMT in these endometrial cancer cells. In addition, we showed that c-Myc directly binds to and represses the promoter of miR-200 miRNAs, and its up-regulation in TAM-treated endometrial cancer cells leads to the down-regulation of miR-200 and eventually to EMT. Collectively, our data suggest that TAM can repress the miR-200 family and induce EMT via the up-regulation of c-Myc in endometrial cancer cells. These findings describe a possible mechanism of TAM-induced EMT in endometrial cancer and provide a potential new therapeutic strategy for it.
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Neoplasias do Endométrio/induzido quimicamente , Transição Epitelial-Mesenquimal/efeitos dos fármacos , MicroRNAs/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-myc/biossíntese , Tamoxifeno/uso terapêutico , Linhagem Celular Tumoral , Feminino , Humanos , MicroRNAs/biossíntese , Tamoxifeno/efeitos adversosRESUMO
Histone deacetylase (HDAC) inhibitors are emerging as a novel class of anti-tumor agents and have manifested the ability to decrease proliferation and increase apoptosis in different cancer cells. A significant number of genes have been identified as potential effectors responsible for the anti-tumor function of HDAC inhibitor. However, the molecular mechanisms of these HDAC inhibitors in this process remain largely undefined. In the current study, we searched for microRNAs (miRs) that were affected by HDAC inhibitor trichostatin (TSA) and investigated their effects in endometrial cancer (EMC) cells. Our data showed that TSA significantly inhibited the growth of EMC cells and induced their apoptosis. Among the miRNAs that altered in the presence of TSA, the miR-106b-93-25 cluster, together with its host gene MCM7, were obviously down-regulated in EMC cells. p21 and BIM, which were identified as target genes of miR-106b-93-25 cluster, increased in TSA treated tumor cells and were responsible for cell cycle arrest and apoptosis. We further identified MYC as a regulator of miR-106b-93-25 cluster and demonstrated its down-regulation in the presence of TSA resulted in the reduction of miR-106b-93-25 cluster and up-regulation of p21 and BIM. More important, we found miR-106b-93-25 cluster was up-regulated in clinical EMC samples in association with the overexpression of MCM7 and MYC and the down-regulation of p21 and BIM. Thus our studies strongly indicated TSA inhibited EMC cell growth and induced cell apoptosis and cell cycle arrest at least partially through the down-regulation of the miR-106b-93-25 cluster and up-regulation of it's target genes p21 and BIM via MYC.
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Apoptose/efeitos dos fármacos , Regulação para Baixo/genética , Neoplasias do Endométrio/genética , Neoplasias do Endométrio/patologia , Ácidos Hidroxâmicos/farmacologia , MicroRNAs/genética , Proteínas Proto-Oncogênicas c-myc/genética , Apoptose/genética , Proteínas Reguladoras de Apoptose/genética , Proteínas Reguladoras de Apoptose/metabolismo , Proteína 11 Semelhante a Bcl-2 , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Inibidor de Quinase Dependente de Ciclina p21/genética , Inibidor de Quinase Dependente de Ciclina p21/metabolismo , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Regulação para Baixo/efeitos dos fármacos , Elementos E-Box/genética , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , MicroRNAs/metabolismo , Componente 7 do Complexo de Manutenção de Minicromossomo , Família Multigênica/genética , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Regiões Promotoras Genéticas/genética , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Proto-Oncogênicas c-myc/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismoRESUMO
The timing of meals has been suggested to play an important role in circadian regulation and metabolic health. Three meals a day is a well-established human feeding habit, which in today's lifestyle may or may not be followed. The aim of this study was to test whether the absence of breakfast or supper significantly affects the circadian system and physiological function. The authors developed a rat model for their daily three meals study, whereby animals were divided into three groups (three meals, TM; no first meal, NF; no last meal, NL) all fed with the same amount of food every day. Rats in the NF group displayed significantly decreased levels of plasma triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and glucose in the activity phase, accompanied by delayed circadian phases of hepatic peripheral clock and downstream metabolic genes. Rats in the NL group showed lower concentration of plasma TC, HDL-C, and glucose in the rest phase, plus reduced adipose tissue accumulation and body weight gain. Real-time polymerase chain reaction (PCR) analysis indicated an attenuated rhythm in the food-entraining pathway, including down-regulated expression of the clock genes Per2, Bmal1, and Rev-erbα, which may further contribute to the delayed and decreased expression of FAS in lipogenesis in this group. Our findings are consistent with the conclusion that the daily first meal determines the circadian phasing of peripheral clocks, such as in the liver, whereas the daily last meal tightly couples to lipid metabolism and adipose tissue accumulation, which suggests differential physiological effects and function of the respective meal timings.
Assuntos
Ritmo Circadiano/fisiologia , Ingestão de Alimentos/fisiologia , Animais , Glicemia , Proteínas CLOCK/genética , Proteínas CLOCK/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/fisiologia , Glucose/metabolismo , Metabolismo dos Lipídeos , Lipídeos/sangue , Fígado/metabolismo , Masculino , Ratos , Ratos WistarRESUMO
The cascade of caspase processing and activation is the core of apoptotic cell signaling. Initiator caspases are activated by adaptor-mediated clustering, which allows the intermolecular autoprocessing of the zymogens in close proximity. Caspase-8, the prototypical initiator critically involved in apoptosis induced by varied extrinsic stimuli, is physiologically recruited via a classical FasL/Fas/FADD pathway. Meanwhile, artificial models of caspase-8 activation have been proposed via inducible dimerization of a heterologous domain fused to the zymogen. Here, we describe the generation of a chimeric protein of caspase-8 and the ligand-binding domain (LBD) of estrogen receptor α (ERα), which dimerizes and undergoes autocleavage upon engagement by the ligand, estradiol. Breast cancer cells expressing this fusion protein exhibit apoptotic cell death in vitro and suppressed tumor growth in xenograft nude mice models in response to the administration of estrogen. Thus, the suicidal caspase-8/ERα-LBD protein, which reverses the mitogenic effects of estrogens, has potential to provide novel approaches to treating estrogen-dependent and -independent breast cancers.
Assuntos
Neoplasias da Mama/metabolismo , Caspase 8/metabolismo , Transformação Celular Neoplásica/metabolismo , Estradiol/farmacologia , Receptor alfa de Estrogênio/metabolismo , Animais , Apoptose/fisiologia , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Caspase 8/genética , Linhagem Celular Tumoral , Transformação Celular Neoplásica/genética , Transformação Celular Neoplásica/patologia , Dimerização , Modelos Animais de Doenças , Estradiol/metabolismo , Receptor alfa de Estrogênio/genética , Feminino , Humanos , Camundongos , Camundongos Nus , Ligação Proteica , Transfecção , Transplante HeterólogoRESUMO
Bisphosphonates (BPs) have a profound effect on bone resorption and are widely used to treat osteoclast-mediated bone diseases. They suppress bone resorption by inhibiting the activity of mature osteoclasts and/or the formation of new osteoclasts. Osteoblasts may be an alternative target for BPs. Periodontal ligament stem cells (PDLSCs) exhibit osteoblast-like features and are capable of differentiating into osteoblasts or cementoblasts. This study aimed to determine the effects of ibandronate, a nitrogen-containing BP, on the proliferation and the differentiation of PDLSCs and to identify the microRNAs (miRNAs) that mediate these effects. The PDLSCs were treated with ibandronate, and cell proliferation was measured using the MTT (3-dimethylthiazol-2,5-diphenyltetrazolium bromide) assay. The expression of genes and miRNAs involved in osteoblastic differentiation was assayed using quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR). Ibandronate promoted the proliferation of PDLSCs and enhanced the expression of alkaline phosphatase (ALP), type I collagen (COL-1), osteoprotegerin (OPG), osteocalcin (OCN), and Runx2. The expression of miRNAs, including miR-18a, miR-133a, miR-141 and miR-19a, was significantly altered in the PDLSCs cultured with ibandronate. In PDLSCs, ibandronate regulates the expression of diverse bone formation-related genes via miRNAs. The exact mechanism underlying the role of ibandronate in osteoblasts has not been completely understood. Ibandronate may suppress the activity of osteoclasts while promoting the proliferation of osteoblasts by regulating the expression of miRNAs.