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Deubiquitinating enzymes (DUBs) regulate antiviral immune response through targeting DNA sensor signaling pathway members. As one of the DNA sensors, interferon (IFN)-γ inducible protein 16 (IFI16) play a major role in response to virus infections through activating the canonical STING/TBK-1/IRF3 signaling pathway. Only a few studies discuss the function of DUBs in IFI16-mediated antiviral response. Ubiquitin-specific protease 12 (USP12), which is one of the major members of the USP family, participates in various biological functions. However, whether USP12 regulates the nucleic acid sensor to modulate antiviral immune responses has not yet been elucidated. In this study, we found that knockout or knockdown of USP12 impaired the HSV-1-induced expressions of IFN-ß, CCL-5, IL-6, and downstream interferon-stimulated genes (ISGs). Moreover, USP12 deficiency increased HSV-1 replication and host susceptibility to HSV-1 infection. Mechanistically, USP12 inhibited the proteasome-dependent degradation of IFI16 through its deubiquitinase activity, thereby maintaining IFI16 stability and promoting IFI16-STING-IRF3- and p65-mediated antiviral signaling. Overall, our findings demonstrate an essential role of USP12 in DNA-sensing signaling and contribute to the understanding of deubiquitination-mediated regulation of innate antiviral responses.
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Herpes Simple , Herpesvirus Humano 1 , Humanos , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Herpesvirus Humano 1/fisiología , Interferones/metabolismo , Antivirales/metabolismo , Inmunidad Innata , Ubiquitina Tiolesterasa/genética , Ubiquitina Tiolesterasa/metabolismo , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismoRESUMEN
[This corrects the article DOI: 10.1371/journal.ppat.1011480.].
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Ubiquitin-specific peptidase 25 (USP25) is one of the best-characterized deubiquitinating enzymes and plays a vital regulatory role in various biological processes, especially in cancer development and immune regulation. However, the exact role of USP25 and its underlying mechanisms in macrophage activation and immunogenicity during Mycobacterium tuberculosis infection remain unclear. In this study, we found that M tuberculosis infection induced USP25 expression in human and mouse macrophages. In particular, USP25 expression is elevated in multiple cell types, especially monocytes, in patients with tuberculosis. Additionally, USP25 deficiency in macrophages and mice resulted in compromised immunity against M tuberculosis infection, accompanied by reduced expressions of various proinflammatory cytokines and chemokines. Mechanistically, USP25 in macrophages promoted the activation of the ERK signaling pathway through deubiquitination and stabilization of B-Raf and C-Raf. These findings collectively suggest the critical roles of USP25 in M tuberculosis infection and its potential as a therapeutic target.
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Daqu is used as the fermentation starter of Baijiu and contributes diversified functional microbes for saccharifying grains and converting sugars into ethanol and aroma components in Baijiu products. Daqu is mainly classified into three types, namely low (LTD), medium (MTD) and high (HTD) temperature Daqu, according to the highest temperatures reached in their fermentation processes. In this study, we used the PacBio small-molecule real-time (SMRT) sequencing technology to determine the full-length 16 S rRNA gene sequences from the metagenomes of 296 samples of different types of Daqu collected from ten provinces in China, and revealed the bacterial diversity at the species level in the Daqu samples. We totally identified 310 bacteria species, including 78 highly abundant species (with a relative abundance >0.1% each) which accounted for 91.90% of the reads from all the Daqu samples. We also recognized the differentially enriched bacterial species in different types of Daqu, and in the Daqu samples with the same type but from different provinces. Specifically, Lactobacillales, Enterobacterales and Bacillaceae were significantly enriched in the LTD, MTD and HTD groups, respectively. The potential co-existence and exclusion relationships among the bacteria species involved in all the Daqu samples and in the LTD, MTD and HTD samples from a specific region were also identified. These results provide a better understanding of the bacterial diversity in different types of Daqu at the species level.
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Bacterias , Fermentación , ARN Ribosómico 16S , ARN Ribosómico 16S/genética , Bacterias/genética , Bacterias/clasificación , Bacterias/aislamiento & purificación , Bacterias/metabolismo , China , Microbiota , Filogenia , ADN Bacteriano/genética , Biodiversidad , Bebidas Alcohólicas/microbiología , Bebidas Alcohólicas/análisis , Microbiología de Alimentos , Metagenoma , Alimentos Fermentados/microbiologíaRESUMEN
Polarization conversion is useful for studies of chiral structures in biology and chemistry, and for polarization diversity in communications. It is conventionally realized with wave plates, which, however, present challenges due to limited material availability, as well as narrow bandwidth and low efficiency at terahertz frequencies. To enhance bandwidth and efficiency, the concept of the Huygens' metasurface is adopted here for a transmissive half-wave plate. The half-wave metasurface is designed following the optimal frequency-independent circuit parameters provided by a broadband semi-analytical approach. Simulation results of an optimal design suggest that a 15-dB extinction ratio can be sustained from 219 GHz to 334 GHz, corresponding to a fractional bandwidth of 41.6%. The measured results indicate that the fabricated structure enables a 15-dB extinction ratio from 220 GHz to 303 GHz, with a cross-polarization transmission efficiency above 76.7% for both linear and circular polarizations. This half-wave metasurface design can be readily integrated into compact terahertz systems for diverse applications.
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This publisher's note contains corrections to Opt. Lett.46, 4164 (2021)OPLEDP0146-959210.1364/OL.431285.
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Far-infrared absorbers exhibiting wideband performance are in great demand in numerous applications, including imaging, detection, and wireless communications. Here, a nonresonant far-infrared absorber with ultra-wideband operation is proposed. This absorber is in the form of inverted pyramidal cavities etched into moderately doped silicon. By means of a wet-etching technique, the crystallinity of silicon restricts the formation of the cavities to a particular shape in an angle that favors impedance matching between lossy silicon and free space. Far-infrared waves incident on this absorber experience multiple reflections on the slanted lossy silicon side walls, being dissipated towards the cavity bottom. The simulation and measurement results confirm that an absorption beyond 90% can be sustained from 1.25 to 5.00 THz. Furthermore, the experiment results suggest that the absorber can operate up to at least 21.00 THz with a specular reflection less than 10% and negligible transmission.
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Based on Boyd's "Observation Orientation-Decision-Action (OODA)" aerial combat theory and the principles of operational success, an analysis of the operational division patterns for cross-generational human-machine collaboration was conducted. The research proposed three stages in the development of aerial combat human-machine fusion intelligence: "Human-Machine Separation, Functional Coordination," "Human-Machine Trust, Task Coordination," and "Human-Machine Integration, Deep Fusion." Currently, the transition from the first stage to the second stage is underway, posing challenges primarily related to the lack of effective methods guiding experimental research on human-machine fusion interaction and trust. Building upon the principles of decision neuroscience and the theory of supply and demand relationships, the study analyzed the decision-making patterns of human-machine fusion intelligence under different states. By investigating the correlations among aerial combat mission demands, dynamic operational limits of human-machine tasks, and aerial combat mission performance, a theoretical model of human-machine fusion interaction and trust was proposed. This model revealed the mechanistic coupling of human-machine interactions in aerial tasks, aiming to optimize the decision-making processes of human-machine systems to enhance mission performance. It provides methodological support for the design and application of intelligent collaborative interaction modes in aviation equipment.
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Descriptive sensory analysis, headspace solid-phase microextraction-gas chromatography-mass spectrometry, gas chromatography-flame ionization detector and multivariate statistical analysis were used to elucidate the regional dependence of sauce-flavor baijiu (SFB). Although SFB samples from different regions couldn't be clearly classified by sensory profiles, they could be clearly divided into 5 groups in principal component analysis plot based on quantitative targeted flavoromics analysis. And then, the relationship between sensory attributes and volatile compounds were investigated by network analysis. Twenty regional aroma markers were identified by multivariate statistical analysis to distinguish SFB samples from different regions. Furthermore, the influence of manufacturing operation on SFB in Guizhou region was further analyzed. Thirty-eight potential compounds were significant different in Guizhou SFB samples with different manufacturing operations. This study not only provides a better understanding of regional dependence on SFB flavor, but also further clarifies the inheritance importance of manufacturing operation in traditional SFB production.
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Compuestos Orgánicos Volátiles , Cromatografía de Gases y Espectrometría de Masas/métodos , Compuestos Orgánicos Volátiles/análisis , Alimentos , Odorantes/análisis , Aromatizantes/análisisRESUMEN
Autoreactive CD4+ T helper cells are critical players that orchestrate the immune response both in multiple sclerosis (MS) and in other neuroinflammatory autoimmune diseases. Ubiquitination is a posttranslational protein modification involved in regulating a variety of cellular processes, including CD4+ T cell differentiation and function. However, only a limited number of E3 ubiquitin ligases have been characterized in terms of their biological functions, particularly in CD4+ T cell differentiation and function. In this study, we found that the RING finger protein 213 (RNF213) specifically promoted regulatory T (Treg) cell differentiation in CD4+ T cells and attenuated autoimmune disease development in an FOXO1-dependent manner. Mechanistically, RNF213 interacts with Forkhead Box Protein O1 (FOXO1) and promotes nuclear translocation of FOXO1 by K63-linked ubiquitination. Notably, RNF213 expression in CD4+ T cells was induced by IFN-ß and exerts a crucial role in the therapeutic efficacy of IFN-ß for MS. Together, our study findings collectively emphasize the pivotal role of RNF213 in modulating adaptive immune responses. RNF213 holds potential as a promising therapeutic target for addressing disorders associated with Treg cells.
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Diferenciación Celular , Proteína Forkhead Box O1 , Interferón beta , Linfocitos T Reguladores , Ubiquitina-Proteína Ligasas , Ubiquitinación , Linfocitos T Reguladores/inmunología , Linfocitos T Reguladores/metabolismo , Ubiquitina-Proteína Ligasas/metabolismo , Ubiquitina-Proteína Ligasas/genética , Animales , Proteína Forkhead Box O1/metabolismo , Proteína Forkhead Box O1/genética , Ratones , Humanos , Interferón beta/metabolismo , Ratones Endogámicos C57BL , Núcleo Celular/metabolismo , Esclerosis Múltiple/inmunología , Esclerosis Múltiple/metabolismo , Esclerosis Múltiple/genética , Esclerosis Múltiple/patología , Transporte Activo de Núcleo Celular , Femenino , Ratones Noqueados , Encefalomielitis Autoinmune Experimental/inmunología , Encefalomielitis Autoinmune Experimental/metabolismo , Encefalomielitis Autoinmune Experimental/genética , Células HEK293RESUMEN
Mechanized, automated and intelligent brewing is an important trend of innovation and transition in Jiang-flavor baijiu industry. In this study, physicochemical parameters, microbial community composition and flavor substances during 3rd round heap fermentation between mechanical and traditional workshop were investigated and compared based on traditional culturable methods, high-throughput sequencing technology and gas chromatography analysis. The dominant bacterial and fungal genera were consistent between the two workshops, but mechanized brewing had a significant impact on the composition of fungal communities. Rhodococcus and Monascus were special genera in mechanical workshop. The interaction relationship between physicochemical parameters and dominant microorganisms in mechanized workshop was different from traditional workshop as well. This study provided a scientific basis for further analyzing the mechanism of mechanized brewing of Jiang-flavor baijiu. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-023-01483-y.
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Background: CD4+ T cells play an important role in body development and homeostasis. Quantitative and functional changes in CD4+ T cells result in abnormal immune responses, which lead to inflammation, cancer, or autoimmune diseases, such as multiple sclerosis (MS). Ubiquitination plays an essential role in the differentiation and functioning of CD4+ T cells. However, the function of several E3 ubiquitin ligases in CD4+ T cell differentiation and T cell-mediated pathological diseases remains unclear. Methods: RNA sequencing data were analyzed to identify the E3 ubiquitin ligases that participate in the pathogenesis of MS. Furthermore, conditional knockout mice were generated. Specifically, flow cytometry, qPCR, western blot, CO-IP and cell transfer adoptive experiments were performed. Results: In this study, we identified The RING finger 157 (RNF157) as a vital regulator of CD4+ T cell differentiation; it promoted Th1 differentiation but attenuated Th17 differentiation and CCR4 and CXCR3 expressions in CD4+ T cells, thereby limiting experimental autoimmune encephalomyelitis development. Mechanistically, RNF157 in CD4+ T cells targeted HDAC1 for K48-linked ubiquitination and degradation. Notably, RNF157 expression was significantly decreased and showed a significant negative correlation with RORγt expression in patients with MS. Conclusions: Our study highlights the critical role of RNF157 in regulating CD4+ T cell functions in autoimmune diseases and suggests RNF157 as a potential target in adaptive immune responses against MS and other autoimmune disorders.
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Encefalomielitis Autoinmune Experimental , Esclerosis Múltiple , Ratones , Animales , Autoinmunidad , Ubiquitinación , Encefalomielitis Autoinmune Experimental/metabolismo , Esclerosis Múltiple/metabolismo , Ubiquitina-Proteína Ligasas/genética , Ubiquitina-Proteína Ligasas/metabolismo , Diferenciación Celular , Ratones Noqueados , Linfocitos T CD4-Positivos , Ubiquitinas/metabolismo , Ratones Endogámicos C57BLRESUMEN
The balance between inflammatory T helper type 17 (Th17) and immunosuppressive regulatory T (Treg) cells is critical for maintaining immune homeostasis in the human body and is tightly regulated under healthy conditions. An increasing number of studies have reported that deubiquitinases (DUBs) play a vital role in regulating Th17- and Treg-cell differentiation. However, the biological functions of only a small fraction of DUBs in Th17- and Treg-cell differentiation are well defined. In this study, we identified ubiquitin-specific peptidase 1 (USP1) as a vital regulator of CD4+ T-cell differentiation. USP1 promoted Th17-cell differentiation but attenuated Treg-cell differentiation, thereby promoting the development of inflammatory diseases. Mechanistically, USP1 in CD4+ T cells enhanced the activity of RORγt but promoted the proteasomal degradation of Foxp3 through deubiquitination and stabilization of TAZ in vitro and in vivo. Notably, ML323, a specific inhibitor of the USP1/UAF1 deubiquitinase complex, inhibited Th17-cell differentiation and promoted Treg-cell differentiation in vitro and in vivo, indicating that ML323 might be a promising candidate for the treatment of diseases associated with an imbalance between Th17 and Treg cells. Our study highlights the critical role of USP1 in regulating adaptive immune responses and suggests that USP1 might be a drug target for the treatment of diseases associated with an imbalance between Th17 and Treg cells.
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Linfocitos T Reguladores , Células Th17 , Humanos , Diferenciación Celular , Factores de Transcripción , Proteasas Ubiquitina-EspecíficasRESUMEN
Ubiquitination is an important protein modification that regulates diverse biological processes, including CD4+ T cell differentiation and functions. However, the function of most E3 ubiquitin ligases in CD4+ T cell differentiation and CD4+ T cell-mediated pathological diseases remains unclear. In this study, we find that tripartite motif-containing motif 11 (TRIM11) specifically negatively regulates regulatory T (Treg) cell differentiation in CD4+ T cells and promotes autoimmune disease development in an AIM2-dependent manner. Mechanistically, TRIM11 interacts with absent in melanoma 2 (AIM2) and promotes the selective autophagic degradation of AIM2 by inducing AIM2 ubiquitination and binding to p62 in CD4+ T cells. AIM2 attenuates AKT and FOXO1 phosphorylation, MYC signaling, and glycolysis, thereby promoting the stability of Treg cells during experimental autoimmune encephalomyelitis (EAE). Our findings suggest that TRIM11 serves as a potential target for immunotherapeutic intervention for dysregulated immune responses that lead to autoimmunity and cancers.
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Encefalomielitis Autoinmune Experimental , Melanoma , Linfocitos T Reguladores , Animales , Diferenciación Celular , Proteínas Proto-Oncogénicas c-myc/metabolismo , Transducción de Señal/fisiología , Linfocitos T Reguladores/metabolismo , Ubiquitina-Proteína Ligasas/metabolismo , Proteína Sequestosoma-1/metabolismo , Autofagia/genética , Autofagia/fisiologíaRESUMEN
Cutaneous squamous cell carcinoma (CSCC) is a widespread malignancy but has a very low long-term survival rate for patients at the metastatic stage. Therefore, it is urgent to identify prognostic biomarkers for CSCC and improve our understanding of disease progression. Here we took advantage of a data-independent acquisition (DIA)-based nano liquid chromatography equipped with an orbitrap mass spectrometry (nLC-MS/MS) and ultraperformance LC coupled to a time-of-flight tandem MS (UPLC-TOF-MS/MS) technique to analyze cancer and corresponding noncancerous tissues from 20 CSCC patients for integrated proteomic and metabolomic analyses. Overall, 6241 tissue proteins were detected, while 136 proteins were significantly expressed in CSCC tissues. Further functional analysis revealed that various biological processes were highly enriched and participated in the pathogenesis of CSCC, especially DNA damage responses. Moreover, 641 named metabolites in total were identified, among which 181 were significantly changed in CSCC tissues. A total of 101 pathways were significantly enriched including apoptosis, autophagy, PI3K-Akt and mTOR signaling pathways. Interestingly, two pathways, protein digestion & absorption and platelet activation were both enriched in proteomic and metabolomic studies involving 5 proteins and 11 metabolites. Accordingly, a four-metabolite panel consisting of arachidonate, glutamine, glutamic acid, and proline (all area under the curve (AUC) values more than 0.9) was developed with a high accuracy (0.971) to distinguish the 20 detected cancer tissues from their noncancerous tissues. Collectively, our work highlighted the key elements and regulatory pathways involved in the pathogenesis of CSCC. More importantly, the present study not only provided potential biomarkers for the early diagnosis of CSCC, but also expanded our knowledge of the physiopathology of the disease. SIGNIFICANCE: CSCC is the second most common human cancer but has few treatment options and few sensitive biomarkers for diagnosis. Here we comprehensively revealed its molecular characteristics by performing integrated tissue proteomic and metabolomic analyses. Significantly distinct profiles and certain enriched pathways including DNA damage responses were identified as associated with CSCC. Moreover, protein digestion & absorption and platelet activation were both enriched in the proteome and metabolome. These identified molecular changes probably play significant roles in CSCC development. Finally, we developed a four-metabolite panel to distinguish CSCC with high accuracy. Overall, our data not only provided potential diagnostic biomarkers, but also extended knowledge on the pathogenesis of CSCC.
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Carcinoma de Células Escamosas , Neoplasias Cutáneas , Humanos , Metaboloma , Fosfatidilinositol 3-Quinasas , Proteoma/metabolismo , Proteómica , Neoplasias Cutáneas/diagnóstico , Espectrometría de Masas en TándemRESUMEN
BACKGROUND: Oral squamous cell carcinoma (OSCC) is associated with high morbidity and ranks sixth among malignancies worldwide. Increasing evidence suggests that microRNAs (miRNAs or miRs) play a critical role in regulating cancer stem cells (CSCs), which drive the proliferation and spread of OSCC. Therefore, based on the alteration of aberrantly expressed miR-495 and homeobox C6 (HOXC6) by Gene Expression Omnibus (GEO) analysis, we subsequently explore the potential effect of miR-495 on the progression of CSCs in OSCC. METHODS: After the isolation of CSCs from the clinical tissue samples of OSCC patients, the expression of miR-495 and HOXC6 was determined, followed by the validation of the relationship between miR-495 and HOXC6. Subsequently, gain- and loss-function approach was performed to detect the role of miR-495 and HOXC6 in cell proliferation, migration, invasion, cell cycle entry, apoptosis, and epithelial-mesenchymal transition (EMT) of CSCs in OSCC, as well as the tumor growth in vivo. RESULTS: HOXC6 was highly expressed while miR-495 was poorly expressed in OSCC. HOXC6 was verified to be a target gene of miR-495, and miR-495 could inhibit the activation of the TGF-ß signaling pathway. CSCs with miR-495 overexpression or HOXC6 silencing exhibited reversed EMT process; reduced abilities of proliferation, migration, and invasion; and promoted cell apoptosis in vitro. Moreover, inhibited tumor growth was observed in vivo after injection with miR-495 agomir or sh-HOXC6. In contrast, the downregulation of miR-495 showed an induced role in the progression of OSCC. CONCLUSION: These findings suggest that miR-495 may suppress HOXC6 to inhibit EMT, proliferation, migration, and invasion while promoting apoptosis of CSCs in OSCC by inhibiting the TGF-ß signaling pathway.
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Carcinoma de Células Escamosas , Neoplasias de Cabeza y Cuello , MicroARNs , Neoplasias de la Boca , Carcinoma de Células Escamosas/genética , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Regulación Neoplásica de la Expresión Génica , Genes Homeobox , Proteínas de Homeodominio/genética , Humanos , MicroARNs/genética , Neoplasias de la Boca/genética , Células Madre Neoplásicas , Transducción de Señal , Carcinoma de Células Escamosas de Cabeza y Cuello , Factor de Crecimiento Transformador beta/genéticaRESUMEN
FZD8, a G protein-coupled receptor protein belonging to the Frizzled family, is considered to play an important role in cancer invasion and metastasis. However, the function of FZD8 in the invasion and metastasis of gastric cancer (GC) has not been elucidated. In this study, we first confirm that FZD8 protein expression was significantly upregulated in gastric cancer tissue and has a potential to be an independent predictor of poor prognosis for patients with GC. In vivo and in vitro evidences were provided that support the idea of FZD8 being able to suppress GC cell invasion and metastasis. Further studies show that FZD8 promotes the markers expression related to invasion and metastasis. FZD8 exerts biological function through the ß-catenin pathway which plays an important role in invasion and metastasis of gastric cancer cells. Finally, FZD8 could activate the ß-catenin pathway and its target gene's expression. In conclusion, our findings show that FZD8 promotes GC invasion and metastasis via the ß-catenin pathway.
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Adenocarcinoma/secundario , Biomarcadores de Tumor/metabolismo , Regulación Neoplásica de la Expresión Génica , Receptores de Superficie Celular/metabolismo , Neoplasias Gástricas/patología , Proteína Wnt1/metabolismo , beta Catenina/metabolismo , Adenocarcinoma/genética , Adenocarcinoma/metabolismo , Biomarcadores de Tumor/genética , Femenino , Estudios de Seguimiento , Humanos , Metástasis Linfática , Masculino , Persona de Mediana Edad , Invasividad Neoplásica , Pronóstico , Receptores de Superficie Celular/genética , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismo , Tasa de Supervivencia , Células Tumorales Cultivadas , Vía de Señalización Wnt , Proteína Wnt1/genética , beta Catenina/genéticaRESUMEN
MicroRNA21 (miR21) is a potential therapeutic target for melanoma. Whether miR21 inhibitor affects the anticancer activity of doxorubicin assisted by c(RGDyK)modified liposome (DLN) in melanoma and the underlying mechanisms are largely unknown. In this study, in vitro and animal models were used to explore the effect of DLN combined with miR21 inhibitor on melanoma cells. The data demonstrated that treatment with 5 µl DLN (final concentration of doxorubicin 5 mg/ml) for 72 h effectively inhibited melanoma cell growth (~75% inhibition). The experiments were then divided into five groups: Control group, vector group, DLN group, miR21 inhibitor group and miR21 inhibitor + DLN group. Compared with the control group, DLN (5 µl) or miR21 inhibitor significantly reduced migration and invasion of melanoma cells, promoted apoptosis and arrested cells at the G1 phase. Notably, the combined application of DLN with miR21 inhibitor further promoted the anticancer effects (reducing migration and invasion of melanoma cells, promoting apoptosis and arresting cells at G1 phase) compared with individual application of DLN or miR21 inhibitor. Mechanically, DLN did not function by reducing miR21 expression, whereas DLN and miR21 inhibitor downregulated Bcell lymphoma-2 (BCL2) expression, and facilitated BCL2associated X protein (Bax) and P53 expression in melanoma cells. DLN and miR21 inhibitor together displayed stronger effects on Bcl2, Bax and P53 expression that each alone. In vivo data further demonstrated that DLN inhibited tumor growth further than a similar dose of doxorubicin only. Furthermore, miR21 inhibitor and DLN exerted the optimal anticancer effect compared with single application of DLN or miR21 inhibitor. Together, the findings demonstrated miR21 inhibitor facilitated the anticancer activity of DLN in melanoma, and the mechanisms involved Bcl2, Bax and P53 expression.
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Doxorrubicina/administración & dosificación , Melanoma Experimental/tratamiento farmacológico , MicroARNs/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Proteína p53 Supresora de Tumor/metabolismo , Proteína X Asociada a bcl-2/metabolismo , Animales , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Terapia Combinada , Doxorrubicina/farmacología , Sinergismo Farmacológico , Femenino , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Liposomas , Melanoma Experimental/genética , Melanoma Experimental/metabolismo , Ratones , MicroARNs/genética , Nanopartículas , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
MicroRNAs (miRNAs) are involved in the maintenance of the cancer stem cell (CSC) phenotype by binding to genes and proteins that modulate cell proliferation and/or cell apoptosis. In our study, we aimed to investigate the role of miR-1305 in the proliferation and self-renewal of liver CSCs (LCSCs) via the ubiquitin-conjugating enzyme E2T (UBE2T)-mediated Akt-signaling pathway. Differentially expressed genes in human hepatocellular carcinoma (HCC) were obtained by in silico analysis. The relationship between miR-1305 and UBE2T was verified by dual luciferase reporter gene assay. qRT-PCR and western blot analysis were performed to determine the expression of UBE2T, the Akt-signaling pathway, and stemness-related factors in LCSCs. In addition, miR-1305 disrupted the activation of the Akt-signaling pathway by targeting UBE2T, and, ultimately, it repressed the sphere formation, colony formation, and proliferation, as well as tumorigenicity of LCSCs. In summary, miR-1305 targeted UBE2T to inhibit the Akt-signaling pathway, thereby suppressing the self-renewal and tumorigenicity of LCSCs. Those findings may provide an enhanced understanding of miR-1305 as a therapeutic target to limit the progression of LCSCs.
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BACKGROUND: This study was conducted to investigate the effects of c[RGDyk]-coated liposomes loaded with Adriamycin (nanodrug) and miR-21 mimics on hepatoma cell line SMCC-7721. METHODS: SMCC-7721 cells were divided into five groups: control (receiving no treatment), nanodrug, miR-21 mimics + nanodrug and miR-21 mimics and empty vector. The concentration and duration of treatments were determined using the MTT assay. Cell apoptosis was detected using flow cytometer. The expression of Bax, Bcl-2 and p53 was measured using qPCR and Western blot analysis. RESULTS: MTT showed that the nanodrug inhibited cell proliferation. Nanodrug and miR-21 led to cell arrest at S phase and apoptosis. qPCR showed that cells treated with nanodrug and miR-21 increased the expression of Bax and p53. Western blot analysis indicated that Bcl-2 expression was significantly reduced. CONCLUSIONS: Our work demonstrates that nanodrug and miR-21 have inhibitory effect on SMCC-7721 cells via up-regulating Bax and p53.