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1.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 52(2): 235-240, 2021 Mar.
Artículo en Chino | MEDLINE | ID: mdl-33829697

RESUMEN

Objective: To explore the influence for combination of Dingkun Dan with estradiol valerateon in treating rats with thin endometrium with Kidney-Yang deficiency based on Wnt/ß-catenin signaling pathway. Methods: The estrous period 40 rats were randomly divided into the normal control group, the model group, the Dingkun Dan group, the estradiol valerateon group and the combination group, with 8 rats in each group. In addition to the normal control group, the rat model of thin endometrium with Kidney-Yang deficiency was established in other groups. The control group used free diet, the model group was given distilled water, the estradiol valerateon group was treated with progynova by gavage at 0.3 mg/(kg·d) , Dingkun Dan group was treated with Dingkun Dan by gavage at 2.26 g/(kg·d), and the combined group was given Dingkun Dan at 2.26 g/(kg·d) on the basis of progynova at 0.3 mg/(kg·d). After 3 estrous cycles, the rats were killed and harvested. HE staining was used to observe histopathologic changes in endometrium. The expression of VEGF in rats endometrium were detected by immunohistochemistry. The expression of ß-catenin, E-cadherin and MMP-9 protein in rat endometrium was detected by Western blot. Results: Compared with the control group, the uterine cavity was narrowed or enlarged, the endometrium glands and blood vessels were sparse, and the endometrium was thinner significantly in the model group ( P<0.01); the levels of VEGF was decreased significantly ( P<0.01), while ß-catenin, E-cadherin and MMP-9 were increased significantly ( P<0.01). Compared with the model group, more endometrial glands, rich intimal vessels, the endometrium were thickened significantly in the 3 treatment groups ( P<0.01 or P<0.05); the levels of VEGF was increased differently. The protein levels of ß-catenin and E-cadherin were significantly decreased in each treatment group ( P<0.01), and MMP-9 were significantly decreased in the Dingkun Dan group and in the combination group ( P<0.01). Compared with the estradiol valerateon group, the level of ß-catenin in Dingkun Dan group was higher, and MMP-9 was lower ( P<0.05 or P<0.01). The levels of ß-catenin and MMP-9 in the combination group were significantly decreased ( P<0.01). Compared with the combination group, the levels of ß-catenin was increased significantly, while decreased signicantly in Dingkun Dan group ( P<0.01 or P<0.05). Conclusion: Dingkun Dan combined with estradiol valerateon can increase the thicken of the endometrium by up-regulation of VEGF, while down-regulate of ß-catenin, E-cadherin and MMP-9 in rats with Shen-Yang deficiency and thin endometrium.


Asunto(s)
Estradiol , Vía de Señalización Wnt , Animales , Endometrio/metabolismo , Femenino , Riñón , Ratas , Deficiencia Yang , beta Catenina/genética , beta Catenina/metabolismo
2.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 29(2): 433-438, 2021 Apr.
Artículo en Chino | MEDLINE | ID: mdl-33812411

RESUMEN

OBJECTIVE: To investigate the significance of low-density lipoprotein receptor-related protein 5 and 6 (LRP5/6) in the Wnt/ß-catenin signaling pathway in the pathogenesis and prognosis of childhood acute lymphoblastic leukemia (ALL). METHODS: A total of 43 children who were newly diagnosed and achieved complete remission after remission induction therapy were enrolled. The children before treatment were included in incipient group, and after treatment when achieved complete remission included in remission group. A total of 39 children with immune thrombocytopenia were enrolled in control group. Three milliliter bone marrow samples were collected from above-mentioned each group. QRT-PCR was used to determine the mRNA expression of LRP5 and LRP6 in blood mononuclear cells of bone marrow. Western blot was used to detect the protein expression of LRP5 and LRP6. According to the protein expression levels of LRP5 and LRP6, the children were divided into low-expression group and high-expression group, and the clinical biological characteristics were compared between these two groups. Survival analysis was performed by Kaplan-Meier method. RESULTS: Both mRNA and protein expression levels of LRP5 and 6 were upregulated in the incipient group compared with the control and remission group (P<0.05). The mRNA and protein expressions of LRP5 and LRP6 in the high-risk group were higher than those in the medium-risk group (P<0.05), it is the same as in the medium-risk group than the low-risk group (P<0.05). The mRNA and protein expressions of LRP5 and 6 positively correlated with risk degree in the incipient group (rLRP5 mRNA=0.84, P<0.05; rLRP6 mRNA=0.66, P<0.05; rLRP5 protein=0.82, P<0.05; rLRP6 protein=0.76, P<0.05). The white blood cell count and lactate dehydrogenase in LRP5 and LRP6 high expression group were significantly higher than those in low expression group (P<0.05), while there was no significant difference in other biological characteristics. Kaplan-meier survival analysis showed that in the 43 children 3-year overall survival rate and event-free survival rate was (91.7±4.7)% and (87.6±5.2)%, respectively. CONCLUSION: The high expression of LRP5/6 may be one of the pathogenesis of childhood ALL, and the degree of LRP5/6 increase may be related to the risk level.


Asunto(s)
Leucemia-Linfoma Linfoblástico de Células Precursoras , Vía de Señalización Wnt , Niño , Humanos , Lipoproteínas LDL , Proteína-5 Relacionada con Receptor de Lipoproteína de Baja Densidad , Receptores de LDL , beta Catenina/metabolismo
3.
Nat Commun ; 12(1): 1368, 2021 03 01.
Artículo en Inglés | MEDLINE | ID: mdl-33649334

RESUMEN

The homeostasis of the gut epithelium relies upon continuous renewal and proliferation of crypt-resident intestinal epithelial stem cells (IESCs). Wnt/ß-catenin signaling is required for IESC maintenance, however, it remains unclear how this pathway selectively governs the identity and proliferative decisions of IESCs. Here, we took advantage of knock-in mice harboring transgenic ß-catenin alleles with mutations that specifically impair the recruitment of N- or C-terminal transcriptional co-factors. We show that C-terminally-recruited transcriptional co-factors of ß-catenin act as all-or-nothing regulators of Wnt-target gene expression. Blocking their interactions with ß-catenin rapidly induces loss of IESCs and intestinal homeostasis. Conversely, N-terminally recruited co-factors fine-tune ß-catenin's transcriptional output to ensure proper self-renewal and proliferative behaviour of IESCs. Impairment of N-terminal interactions triggers transient hyperproliferation of IESCs, eventually resulting in exhaustion of the self-renewing stem cell pool. IESC mis-differentiation, accompanied by unfolded protein response stress and immune infiltration, results in a process resembling aberrant "villisation" of intestinal crypts. Our data suggest that IESC-specific Wnt/ß-catenin output requires selective modulation of gene expression by transcriptional co-factors.


Asunto(s)
Mucosa Intestinal/citología , Células Madre/metabolismo , Factores de Transcripción/metabolismo , Transcripción Genética , beta Catenina/química , beta Catenina/metabolismo , Algoritmos , Animales , Secuencia de Bases , Diferenciación Celular , Proliferación Celular , Cromatina/metabolismo , Ensamble y Desensamble de Cromatina , Homeostasis , Hiperplasia , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Ratones , Proteínas Mutantes/metabolismo , Mutación/genética , Organoides/metabolismo , Fenotipo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Transducción de Señal
4.
Hua Xi Kou Qiang Yi Xue Za Zhi ; 39(1): 74-80, 2021 Feb 01.
Artículo en Inglés, Chino | MEDLINE | ID: mdl-33723940

RESUMEN

OBJECTIVES: This study aimed to explore the effect of sex determining region Y-box 9 (SOX9) on the microtubule formation and epithelial-mesenchymal transition (EMT) of human oral squamous cell carcinoma (OSCC) CAL27 and the underlying mechanism. METHODS: SOX9-shRNA1 and SOX9-shRNA2 were designed and synthesized and then transfected into CAL27 cells. The expression of SOX9 was detected by quantitative real-time polymerase chain reaction. Microtubule formation assay was used to detect the change in the number of microtubule nodules after interfering with SOX9. Immunofluorescence was used to detect the Vimentin content. Western blot was used to detect the protein expression of EMT marker molecules and Wnt/ß-catenin pathway-related proteins, such as E-cadherin, N-cadherin, Fibronectin, Wnt, ß-catenin, T-cell factor-4 (TCF-4). RESULTS: The expression level of SOX9 significantly decreased after transfection with SOX9-shRNA1 and SOX9-shRNA2 in CAL27 cells (F=578.000, P=0.000; F=96.850, P=0.000). Interference with SOX9 inhibited the EMT of OSCC. After interference with SOX9, the number of tubules and Vimentin positive cells decreased significantly (F=169.700, P=0.000). The expression level of E-cadherin significantly increased (F=181.400, P=0.000). The expression levels of N-cadherin, Fibronectin, Wnt, ß-catenin, and TCF-4 proteins significantly decreased (N-cadherin: F=101.400, P=0.000; Fibronectin: F=122.300, P=0.000; Wnt: F=70.290, P=0.000; ß-catenin: F=81.740, P=0.000; TCF-4: F=37.020, P=0.000). CONCLUSIONS: Interference with SOX9 decreased Vimentin content and inhibited the microtubule formation and protein expression of EMT marker molecules, as well as the expression of proteins related to the Wnt/ß-catenin pathway. Thus, SOX9 can induce microtubule formation and EMT in CAL27, which was related to the inhibition of the Wnt/ß-catenin pathway activation.


Asunto(s)
Carcinoma de Células Escamosas , Neoplasias de Cabeza y Cuello , Neoplasias de la Boca , Factor de Transcripción SOX9 , Línea Celular Tumoral , Transición Epitelial-Mesenquimal , Humanos , Microtúbulos/metabolismo , Factor de Transcripción SOX9/metabolismo , Carcinoma de Células Escamosas de Cabeza y Cuello , Vía de Señalización Wnt , beta Catenina/metabolismo
5.
Chem Biol Interact ; 337: 109366, 2021 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-33549581

RESUMEN

Tripartite motif-containing protein 26 (TRIM26) is a member of the TRIM protein family and has been demonstrated to play crucial roles in several types of cancers. However, the biological role of TRIM26 in bladder cancer and the mechanism have not been studied. In this study, we investigated the expression of TRIM26 in bladder cancer tissues and their adjacent non-tumor tissues by Western blot and qRT-PCR. In vitro investigations were performed to assess the roles of TRIM26 in bladder cancer using TRIM26-silencing and TRIM26-overexpressing bladder cancer cell lines. MTT and EdU assays were performed to evaluate cell proliferation. Cell migration and invasion were determined by transwell assays. Western blot analysis was performed to detect the expression levels of p-Akt, Akt, p-GSK3ß, GSK3ß, ß-catenin and c-Myc. Our results showed that TRIM26 expression was upregulated in human bladder cancer tissues and cell lines at both mRNA and protein levels. Knockdown of TRIM26 significantly inhibited the proliferation, migration and invasion of bladder cancer cells. In contrast, TRIM26 overexpression promoted bladder cancer cell proliferation, cell migration and invasion. Furthermore, knockdown of TRIM26 significantly decreased the levels of p-Akt, p-GSK3ß, ß-catenin and c-Myc in bladder cancer cells. Additionally, induction of Akt by SC79 treatment reversed the inhibitory effects of TRIM26 knockdown on the cellular behaviors of bladder cancer cells, while inhibition of ß-catenin reversed the effects of TRIM26 overexpression on the behaviors. Finally, knockdown of TRIM26 attenuated the growth of tumor xenografts in nude mice. In conclusion, these findings demonstrated that TRIM26 exerted an oncogenic role in bladder cancer through regulation of cell proliferation, migration and invasion via the Akt/GSK3ß/ß-catenin pathway.


Asunto(s)
Proliferación Celular , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Proteínas de Motivos Tripartitos/metabolismo , Ubiquitina-Proteína Ligasas/metabolismo , Neoplasias de la Vejiga Urinaria/patología , beta Catenina/metabolismo , Acetatos/farmacología , Animales , Benzopiranos/farmacología , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Humanos , Ratones , Ratones Desnudos , Proteínas Proto-Oncogénicas c-akt/agonistas , Interferencia de ARN , ARN Interferente Pequeño/metabolismo , ARN Interferente Pequeño/uso terapéutico , Transducción de Señal/efectos de los fármacos , Trasplante Heterólogo , Proteínas de Motivos Tripartitos/antagonistas & inhibidores , Proteínas de Motivos Tripartitos/genética , Ubiquitina-Proteína Ligasas/antagonistas & inhibidores , Ubiquitina-Proteína Ligasas/genética , Neoplasias de la Vejiga Urinaria/tratamiento farmacológico , Neoplasias de la Vejiga Urinaria/metabolismo
6.
Life Sci ; 272: 119208, 2021 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-33582177

RESUMEN

AIMS: The efficacy of anti-osteoporotic treatments is still limited. Our study aimed to investigate the effect of extracellular vesicles (EVs) derived from bone marrow-derived MSCs (BMSCs) overexpressing glycoprotein non-melanoma clone B (GPNMB) on osteoporosis (OP). MAIN METHODS: Lentiviral vector for GPNMB overexpression or its negative control was generated and transfected into BMSCs. EVs enriched with GPNMB (GPNMB-EVs) were extracted from GPNMB-modified BMSC-conditioned medium and then identified. Cellular uptake and proliferation were analyzed using the Dil-labeled assay and CCK-8 assay, respectively. Cytochemical staining, western blot, and RT-qPCR analysis were performed to assess the effect of GPNMB-EVs on osteogenic differentiation of BMSCs in vitro. Dickkopf-1 (DKK1) as the inhibitor was applied to explore the Wnt/ß-catenin signaling pathway involved in the GPNMB-EV-induced osteogenic differentiation. In vivo experiments were conducted using an ovariectomized (OVX) rat model of postmenopausal osteoporosis, and then assessed the effect of GPNMB-EVs by micro-CT, and histological and immunohistochemical assays. KEY FINDINGS: GPNMB-EVs were taken up by BMSCs, and they noticeably promoted the proliferation of BMSCs. Additionally, GPNMB-EVs activated the Wnt/ß-catenin signaling to stimulate osteogenesis in BMSCs. In vivo examination showed that GPNMB-EVs remarkably improved trabecular bone regeneration and alleviated the osteoporotic phenotype in the OVX-induced rat model of OP. SIGNIFICANCE: EVs derived from GPNMB-modified BMSCs significantly stimulated the proliferation and osteogenic differentiation of BMSCs via the activation of Wnt/ß-catenin signaling and attenuated the bone loss in the OVX-induced rat model of OP. Our findings suggest the promising potential of GPNMB-EVs as cell-free therapy for the treatment of OP.


Asunto(s)
Vesículas Extracelulares/metabolismo , Glicoproteínas de Membrana/farmacología , Osteoblastos/metabolismo , Animales , Médula Ósea/metabolismo , Células de la Médula Ósea/metabolismo , Huesos/metabolismo , Diferenciación Celular/fisiología , Proliferación Celular/fisiología , Células Cultivadas , Femenino , Glicoproteínas de Membrana/metabolismo , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Osteoblastos/fisiología , Osteogénesis/efectos de los fármacos , Osteoporosis/metabolismo , Ovariectomía , Ratas , Vía de Señalización Wnt/efectos de los fármacos , beta Catenina/metabolismo
7.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 37(2): 132-139, 2021 Feb.
Artículo en Chino | MEDLINE | ID: mdl-33504419

RESUMEN

Objective To investigate the effects of the interaction between Wnt/ß-catenin signal and NADPH oxidase 4 (NOX4) on the proliferation of pulmonary epithelial cells in response to silica exposure. Methods The airway instillation of silica to C57BL/6 mice was used to produce mouse silicosis models. Immunohistochemistry was used to determine NOX4 expression in the lungs of silicosis mice. Human lung epithelial cells (BEAS-2B) were exposed to silica to generate an oxidative injury epithelial cell model in vitro. Wnt signal conditioned medium (Wnt3a-CM) and Wnt signal inhibitor XAV939 were used to alter the activity of Wnt signal. An infection adenoviral vector expressing short hairpin RNA to NOX4 (NOX4 shRNA) was used to knock down NOX4 expression in BEAS-2B cells. Western blotting was performed to access the expression of Wnt3a, active-ß-catenin (ABC), transcription factor 4 (TCF4), cyclin D1 and NOX4 proteins in lung tissues and human lung epithelial cells. CCK-8 assay was used to determine the effects of silica of different concentrations on cell viability as well as the effects of NOX4 expression knockdown on cell proliferation in human lung epithelial cells. CellROXR fluorescent probe loading assay was used to detect the release of reactive oxygen species (ROS). Results Mouse silicosis model and BEAS-2B cell model of oxidative damage were successfully generated. The stimulation of silica significantly activated Wnt/ß-catenin signal and induced NOX4 expression, sequentially resulting in ROS production. While ROS scavenger N-acetyl-L-cysteine (NAC) inhibited the silica-induced release of ROS, and then inhibited the expression of ABC protein and Wnt/ß-catenin signal activity. An activation of Wnt signaling induced by Wnt3a-conditioned medium (Wnt3a-CM) increased the expression of NOX4, whereas the Wnt signal inhibitor XAV939 inhibited the expression of NOX4. The expression of Wnt/ß-catenin signal ABC and cyclin D1 and cell proliferation were significantly inhibited by the shRNA-mediated suppression of NOX4 expression. Conclusion Blocking Wnt/-catenin signal and down-regulating NOX4 expression inhibit the proliferation of lung epithelial cells and the damage repair of lung epithelial cells induced by the silica exposure.


Asunto(s)
Dióxido de Silicio , beta Catenina , Animales , Células Epiteliales/metabolismo , Pulmón/metabolismo , Ratones , Ratones Endogámicos C57BL , NADPH Oxidasa 4/genética , Dióxido de Silicio/toxicidad , Vía de Señalización Wnt , beta Catenina/genética , beta Catenina/metabolismo
8.
Bratisl Lek Listy ; 122(2): 132-137, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33502882

RESUMEN

OBJECTIVES: We aimed to investigate the effects of juglone on angiogenesis, metastasis and cell proliferation processes in pancreatic cancer  (PC)  and to understand whether its possible effects occur via the Wnt signaling pathway by analyzing the expression levels of target genes of Wnt signaling. BACKGROUND: PC is a silent and lethal cancer type which can only be detectable after metastasis and angiogenesis processes occured. The Wnt signaling pathway is one of the pathways that plays an active role in many biological processes in the cell. Mutations in the genes of this signaling pathway are related to the development of many cancers. Juglone, a natural compound, is shown to have cytotoxic and apoptotic effects on various cancer cells. METHODS: PANC-1 and BxPC-3 pancreatic cancer cells were treated with juglone at

Asunto(s)
Neoplasias Pancreáticas , Línea Celular Tumoral , Proliferación Celular , Regulación Neoplásica de la Expresión Génica , Humanos , Naftoquinonas , Neovascularización Patológica , Neoplasias Pancreáticas/tratamiento farmacológico , Neoplasias Pancreáticas/genética , Vía de Señalización Wnt , beta Catenina/genética , beta Catenina/metabolismo
9.
Life Sci ; 270: 119072, 2021 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-33482187

RESUMEN

BACKGROUND: Accumulating evidence has reported the role of microRNA (miR) on ischemic brain injury. We aim to investigate the mechanism of miR-376b-5p/Sex-determining region Y-box 7 (SOX7)/Wnt/ß-catenin axis in mice with ischemic brain injury. METHODS: Transient middle cerebral artery occlusion (tMCAO) model was established by suture method. Expression levels of miR-376b-5p, SOX7, and Wnt/ß-catenin pathway-related proteins (Wnt3a and ß-catenin) in brain tissues of tMCAO mice were determined by RT-qPCR and western blot analysis. The target relationship between miR-376b-5p and SOX7 was tested by bioinformatics analysis and luciferase activity assay. The neurological scores of mice were recorded and their behaviors were observed. Moreover, the brain damage, oxidative stress indices, hemoglobin (Hb) content, content of brain water, infarct area, TUNEL positive cells, blood-brain barrier permeability and the number of intact neurons in the ischemic-side brain tissues of tMCAO mice were detected via upregulated miR-376b-5p or downregulated SOX7. RESULTS: In mice with ischemic brain injury, miR-376b-5p targeted and downregulated SOX7 in brain tissues, thus activating the Wnt/ß-catenin pathway. The tMCAO mice showed higher neurological scores, severe brain damage, decreased number of neurons, as well as elevated blood-brain barrier permeability, Hb content, cerebral edema and infarct area. These symptoms could be alleviated by miR-376b-5p elevation or SOX7 inhibition. SOX7 overexpression reversed the effects of miR-376b-5p elevation on tMCAO mice. CONCLUSION: Our study suggests that miR-376b-5p could improve the blood-brain barrier permeability, relieve brain edema and decrease infarct area, thus improve ischemic brain injury via the inhibition of SOX7 and activation of Wnt/ß-catenin pathway.


Asunto(s)
Lesiones Encefálicas/genética , Isquemia Encefálica/genética , MicroARNs/genética , Factores de Transcripción SOXF/metabolismo , Vía de Señalización Wnt , Animales , Lesiones Encefálicas/metabolismo , Isquemia Encefálica/metabolismo , Línea Celular Tumoral , Movimiento Celular/fisiología , Proliferación Celular/fisiología , Modelos Animales de Enfermedad , Infarto de la Arteria Cerebral Media/genética , Infarto de la Arteria Cerebral Media/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , MicroARNs/metabolismo , Factores de Transcripción SOXF/genética , Proteínas Wnt/metabolismo , beta Catenina/metabolismo
10.
Nat Commun ; 12(1): 262, 2021 01 11.
Artículo en Inglés | MEDLINE | ID: mdl-33431859

RESUMEN

The pathogenesis of ulcerative colitis (UC), a major type of inflammatory bowel disease, remains unknown. No model exists that adequately recapitulates the complexity of clinical UC. Here, we take advantage of induced pluripotent stem cells (iPSCs) to develop an induced human UC-derived organoid (iHUCO) model and compared it with the induced human normal organoid model (iHNO). Notably, iHUCOs recapitulated histological and functional features of primary colitic tissues, including the absence of acidic mucus secretion and aberrant adherens junctions in the epithelial barrier both in vitro and in vivo. We demonstrate that the CXCL8/CXCR1 axis was overexpressed in iHUCO but not in iHNO. As proof-of-principle, we show that inhibition of CXCL8 receptor by the small-molecule non-competitive inhibitor repertaxin attenuated the progression of UC phenotypes in vitro and in vivo. This patient-derived organoid model, containing both epithelial and stromal compartments, will generate new insights into the underlying pathogenesis of UC while offering opportunities to tailor interventions to the individual patient.


Asunto(s)
Colitis Ulcerosa/patología , Organoides/patología , Uniones Adherentes/metabolismo , Cadherinas/metabolismo , Progresión de la Enfermedad , Epitelio/patología , Fibroblastos/patología , Humanos , Inflamación/patología , Epiplón/trasplante , Fenotipo , Análisis de Componente Principal , Análisis de Secuencia de ARN , Sulfonamidas/farmacología , Transcriptoma/genética , beta Catenina/metabolismo
11.
Nat Cell Biol ; 23(1): 23-31, 2021 01.
Artículo en Inglés | MEDLINE | ID: mdl-33398177

RESUMEN

A detailed understanding of intestinal stem cell (ISC) self-renewal and differentiation is required to treat chronic intestinal diseases. However, the different models of ISC lineage hierarchy1-6 and segregation7-12 are subject to debate. Here, we have discovered non-canonical Wnt/planar cell polarity (PCP)-activated ISCs that are primed towards the enteroendocrine or Paneth cell lineage. Strikingly, integration of time-resolved lineage labelling with single-cell gene expression analysis revealed that both lineages are directly recruited from ISCs via unipotent transition states, challenging the existence of formerly predicted bi- or multipotent secretory progenitors7-12. Transitory cells that mature into Paneth cells are quiescent and express both stem cell and secretory lineage genes, indicating that these cells are the previously described Lgr5+ label-retaining cells7. Finally, Wnt/PCP-activated Lgr5+ ISCs are molecularly indistinguishable from Wnt/ß-catenin-activated Lgr5+ ISCs, suggesting that lineage priming and cell-cycle exit is triggered at the post-transcriptional level by polarity cues and a switch from canonical to non-canonical Wnt/PCP signalling. Taken together, we redefine the mechanisms underlying ISC lineage hierarchy and identify the Wnt/PCP pathway as a new niche signal preceding lateral inhibition in ISC lineage priming and segregation.


Asunto(s)
Linaje de la Célula , Polaridad Celular , Células Enteroendocrinas/citología , Mucosa Intestinal/citología , Células de Paneth/citología , Células Madre/citología , Proteínas Wnt/metabolismo , Animales , Autorrenovación de las Células , Células Enteroendocrinas/metabolismo , Femenino , Perfilación de la Expresión Génica , Mucosa Intestinal/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Células de Paneth/metabolismo , Receptores Acoplados a Proteínas G/fisiología , Análisis de la Célula Individual , Células Madre/metabolismo , beta Catenina/metabolismo
12.
J Cancer Res Clin Oncol ; 147(4): 1101-1113, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-33471184

RESUMEN

PURPOSE: Recent clinical trials with agents targeting immune checkpoint pathway have emerged as an important therapeutic approach for a broad range of cancer types. Resveratrol has been shown to possess cancer preventive and therapeutic effects and has potential to be chemotherapeutic agent/adjuvant. Here, we assessed the effect of resveratrol on immune checkpoint pathways. METHODS: The expression patterns of Wnt components and PD-L1 were examined by Western blot, Chromatin immunoprecipitation (ChIP) was used for analysis of DNA-protein interaction, the promoter activity was determined by luciferase reporter assay, apoptosis was analyzed by flow cytometry and the ability of the resveratrol to modulate T cell function was assessed in a co-culture system. RESULTS: Although the dose-, and cell-type dependent effects of resveratrol on PD-L1 expression have been reported, we show here that resveratrol dose-dependently upregulates PD-L1 expression at the range of pharmacologic-achievable concentrations in lung cancer cells and that is essential for suppression of T-cell-mediated immune response. We also found that Wnt pathway is critical for mediating resveratrol-induced PD-L1 upregulation. Mechanistically, resveratrol activates SirT1 deacetylase to deacetylate and stabilize transcriptional factor Snail. Snail in turn inhibits transcription of Axin2, which leads in disassembly of destruction complex and enhanced binding of ß-catenin/TCF to PD-L1 promoter. CONCLUSION: We conclude that resveratrol is capable to suppress anti-tumor immunity by controlling mainly PD-L1 expression. This finding will extend the understanding of resveratrol in regulation of tumor immunity and is relevant to the debate on resveratrol supplements for lung cancer patients.


Asunto(s)
Antígeno B7-H1/metabolismo , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Neoplasias Pulmonares/tratamiento farmacológico , Resveratrol/farmacología , Factores de Transcripción de la Familia Snail/metabolismo , Proteínas Wnt/metabolismo , beta Catenina/metabolismo , Antioxidantes/farmacología , Apoptosis , Antígeno B7-H1/genética , Biomarcadores de Tumor/genética , Biomarcadores de Tumor/metabolismo , Proliferación Celular , Humanos , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Factores de Transcripción de la Familia Snail/genética , Células Tumorales Cultivadas , Proteínas Wnt/genética , beta Catenina/genética
13.
J Nat Med ; 75(2): 326-338, 2021 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-33417145

RESUMEN

A methanol extract from Isodonis Herba demonstrated significant proliferative effect on human hair follicle dermal papilla cells (HFDPC, % of control: 150.0 ± 2.0% at 20 µg/mL, p < 0.01). From the extract, 14 ent-kaurane-type diterpenoids (1-14), two abietane-type diterpenoids (15 and 16) and four triterpenoids (17-20) were isolated. Among the isolates, enmein (1, 160.9 ± 3.0% at 20 µM, p < 0.01), isodocarpin (2, 169.3 ± 4.9% at 5 µM, p < 0.01), nodosin (4, 160.5 ± 12.4% at 20 µM, p < 0.01), and oridonin (8, 165.4 ± 10.6% at 10 µM, p < 0.01) showed the proliferative effects. The principal component enmein (1) activated the expression of vascular endothelial growth factor (VEGF) mRNA, upregulated the production of VEGF and increased levels of phospho-Akt, phospho-GSK-3ß, and ß-catenin accumulation in HFDPC, which could be the mechanism of these activate proliferation activity.


Asunto(s)
Diterpenos de Tipo Kaurano/metabolismo , Folículo Piloso/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , beta Catenina/metabolismo , Proliferación Celular , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Humanos
14.
Life Sci ; 269: 118994, 2021 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-33417952

RESUMEN

AIM: To study the role of PARP-1 in EMT of non-small cell lung carcinoma. MATERIALS AND METHODS: We used H1299 and H460 lung cancer cells for knockdown study of PARP-1 using shPARP-1 lentiviral particle. We performed western blotting, confocal microscopy, semi-quantitative PCR, wound healing and colony formation assays. BACKGROUND AND KEY FINDINGS: PARP-1 (poly-ADP ribose polymerase-1) is a multi-domain protein having DNA binding, auto-modification and catalytic domain, that participates in many biological processes including DNA damage detection and repair, transcription regulation, apoptosis, necrosis, cancer progression and metastasis. Metastasis is a leading cause of death in cancer patients, which starts in epithelial tumors via initiating epithelial to mesenchymal transition. There are various transcription factors involved in EMT including Snail-1, Smads, p65, ZEB1 and Twist1. We studied the effect of PARP-1 knockdown on EMT in non-small cell lung cancer cell line H1299. We found a significant increase in epithelial marker including ZO1 and ß-catenin, while prominent decrease in the mesenchymal marker vimentin after PARP-1 knockdown in H1299 cells. Transcription factors including p65, Smad4 and ZEB1 showed significant decrease with concurrent expression of EMT markers. Cell migration and colony formation decreased after PARP-1 knockdown in H1299 cells. SIGNIFICANCE: Overall, the shRNA mediated knockdown of PARP-1 in H1299 cells resulted in reversal of EMT or mesenchymal to epithelial transition (MET) characterized by an increase in epithelial markers and a decrease in mesenchymal markers, via down-regulating transcription factors including Smad4, p65 and ZEB1. Thus PARP-1 has a role in EMT in lung cancer.


Asunto(s)
Carcinoma de Pulmón de Células no Pequeñas/patología , Transición Epitelial-Mesenquimal , Neoplasias Pulmonares/patología , Poli(ADP-Ribosa) Polimerasa-1/metabolismo , Proteína Smad4/metabolismo , Factor de Transcripción ReIA/metabolismo , Homeobox 1 de Unión a la E-Box con Dedos de Zinc/metabolismo , Biomarcadores de Tumor/metabolismo , Carcinoma de Pulmón de Células no Pequeñas/genética , Línea Celular Tumoral , Membrana Celular/metabolismo , Movimiento Celular/genética , Regulación hacia Abajo/genética , Transición Epitelial-Mesenquimal/genética , Regulación Neoplásica de la Expresión Génica , Humanos , Lentivirus/metabolismo , Neoplasias Pulmonares/genética , Modelos Biológicos , ARN Mensajero/genética , ARN Mensajero/metabolismo , ARN Interferente Pequeño/metabolismo , Proteínas de Uniones Estrechas/metabolismo , Ensayo de Tumor de Célula Madre , beta Catenina/metabolismo
15.
Nat Commun ; 12(1): 270, 2021 01 11.
Artículo en Inglés | MEDLINE | ID: mdl-33431892

RESUMEN

Intestinal stem cells (ISCs) residing in the crypts are critical for the continual self-renewal and rapid recovery of the intestinal epithelium. The regulatory mechanism of ISCs is not fully understood. Here we report that CREPT, a recently identified tumor-promoting protein, is required for the maintenance of murine ISCs. CREPT is preferably expressed in the crypts but not in the villi. Deletion of CREPT in the intestinal epithelium of mice (Vil-CREPTKO) results in lower body weight and slow migration of epithelial cells in the intestine. Vil-CREPTKO intestine fails to regenerate after X-ray irradiation and dextran sulfate sodium (DSS) treatment. Accordingly, the deletion of CREPT decreases the expression of genes related to the proliferation and differentiation of ISCs and reduces Lgr5+ cell numbers at homeostasis. We identify that CREPT deficiency downregulates Wnt signaling by impairing ß-catenin accumulation in the nucleus of the crypt cells during regeneration. Our study provides a previously undefined regulator of ISCs.


Asunto(s)
Proteínas de Ciclo Celular/metabolismo , Intestinos/fisiología , Proteínas de Neoplasias/metabolismo , Regeneración/fisiología , Células Madre/metabolismo , Animales , Recuento de Células , Proteínas de Ciclo Celular/deficiencia , Diferenciación Celular , Proliferación Celular , Epitelio/metabolismo , Eliminación de Gen , Humanos , Ratones Endogámicos C57BL , Ratones Noqueados , Modelos Biológicos , Proteínas de Neoplasias/deficiencia , Organoides/metabolismo , Células Madre/citología , Vía de Señalización Wnt , Rayos X , beta Catenina/metabolismo
16.
Am J Surg Pathol ; 45(1): 68-76, 2021 01.
Artículo en Inglés | MEDLINE | ID: mdl-32769429

RESUMEN

CTNNB1 mutations and aberrant ß-catenin expression have adverse prognosis in endometrial endometrioid carcinoma, and recent evidence suggests a prognostic role of ß-catenin in ovarian endometrioid carcinoma. Thus, we aimed to determine the prognostic value of the CTNNB1 mutational status, and its correlation with ß-catenin expression, in a well-annotated cohort of 51 ovarian endometrioid carcinomas. We performed immunohistochemistry for ß-catenin and developed an 11-gene next-generation sequencing panel that included whole exome sequencing of CTNNB1 and TP53. Results were correlated with clinicopathologic variables including disease-free and disease-specific survival. Tumor recurrence was documented in 14 patients (27%), and cancer-related death in 8 patients (16%). CTNNB1 mutations were found in 22 cases (43%), and nuclear ß-catenin in 26 cases (51%). CTNNB1 mutation highly correlated with nuclear ß-catenin (P<0.05). Mutated CTNNB1 status was statistically associated with better disease-free survival (P=0.04, log-rank test) and approached significance for better disease-specific survival (P=0.07). It also correlated with earlier International Federation of Gynecology and Obstetrics stage (P<0.05). Nuclear ß-catenin, TP53 mutations, age, ProMisE group, surface involvement, tumor grade and stage also correlated with disease-free survival. There was no association between membranous ß-catenin expression and disease-free or disease-specific survival. CTNNB1 mutations and nuclear ß-catenin expression are associated with better progression-free survival in patients with OEC. This relationship may be in part due to a trend of CTNNB1-mutated tumors to present at early stage. ß-catenin immunohistochemistry may serve as a prognostic biomarker and a surrogate for CTNN1B mutations in the evaluation of patients with ovarian endometrioid neoplasia, particularly those in reproductive-age or found incidentally without upfront staging surgery.


Asunto(s)
Biomarcadores de Tumor/análisis , Carcinoma Endometrioide , Neoplasias Ováricas , beta Catenina/genética , beta Catenina/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Carcinoma Endometrioide/genética , Carcinoma Endometrioide/metabolismo , Carcinoma Endometrioide/mortalidad , Supervivencia sin Enfermedad , Femenino , Humanos , Persona de Mediana Edad , Mutación , Neoplasias Ováricas/genética , Neoplasias Ováricas/metabolismo , Neoplasias Ováricas/mortalidad , Pronóstico , Resultado del Tratamiento , beta Catenina/análisis
17.
Life Sci ; 267: 118921, 2021 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-33358913

RESUMEN

AIMS: Helicobacter pylori (Hp) infection plays an important role in the development of gastric cancer. Hp can secrete gamma-glutamyltransferase (GGT), however, the impact of GGT of Hp on the human gastric cells is not clear. Although it has been demonstrated that ten-eleven translocation 1 (TET1) is overexpressed in gastric cancer, the relationship between the GGT of Hp and TET1 has not been studied. The aim of this study was to explore the relationship between GGT and TET1, and the role of TET1 in the development of gastric cancer induced by Hp was also explored. MATERIALS AND METHODS: The correlation between TET1 and prognosis of gastric adenoma cancer was analyzed by bioinformatics. The GGT gene of Hp26695 was knocked out by electroporation with plasmid to construct the GGT knockout strain of Hp (Hp-KS-1). The shTET1 lentivirus transfected GES-1, MGC-803 and SGC-7901 cell lines were constructed. The biological characteristics of the three kind of cells were detected. KEY FINDINGS: TET1 was overexpressed in gastric tissues of Hp infected patients and mice. Bioinformatics analysis showed that in patients with gastric cancer, higher TET1 expression would result in poorer prognosis. The GGT gene of Hp can lead to overexpression of TET1 in GES-1, MGC-803 and SGC-7901 cells, along with the activation of Wnt/ß-catenin signaling pathway, and then promoting tumorigenesis. After silencing TET1, the Wnt/ß-catenin signaling pathway which was activated by GGT of Hp was inhibited. SIGNIFICANCE: GGT of Helicobacter pylori can promote gastric carcinogenesis by activating Wnt/ß-catenin signaling pathway trough up-regulating TET1.


Asunto(s)
Infecciones por Helicobacter/microbiología , Helicobacter pylori/enzimología , Helicobacter pylori/genética , Oxigenasas de Función Mixta/metabolismo , Proteínas Proto-Oncogénicas/metabolismo , Neoplasias Gástricas/microbiología , Vía de Señalización Wnt , gamma-Glutamiltransferasa/genética , Pólipos Adenomatosos/metabolismo , Animales , Línea Celular Tumoral , Movimiento Celular/fisiología , Proliferación Celular/fisiología , Transformación Celular Neoplásica , Mucosa Gástrica/metabolismo , Infecciones por Helicobacter/metabolismo , Infecciones por Helicobacter/patología , Helicobacter pylori/metabolismo , Humanos , Masculino , Ratones , Ratones Desnudos , Oxigenasas de Función Mixta/genética , Proteínas Proto-Oncogénicas/genética , Neoplasias Gástricas/metabolismo , Activación Transcripcional , beta Catenina/metabolismo , gamma-Glutamiltransferasa/metabolismo
18.
Arch Biochem Biophys ; 698: 108722, 2021 02 15.
Artículo en Inglés | MEDLINE | ID: mdl-33321112

RESUMEN

ß-Catenin, a key transcriptional factor involved in the canonical Wnt signaling pathway, is regulated by a cascade of phosphorylations and plays a major role in the progression of triple-negative breast cancer (TNBC). However, the phosphorylation induced conformational changes in a ß-Catenin is still poorly understood. Hence, we adopted a conventional molecular dynamics approach to study phosphorylations present in a sequence motif Ser 552 675 and Tyr670 of the ß-Catenin domain and analyzed in terms of structural transitions, bond formation, and folding-misfolding conformations. Our results unveil the ß-Catenin linear motif 549-555 (RRTSMGG) of armadillo repeats domain prefers order to disorder state. In contrast, helix C associated with 670-678 (YKKRLSVEL) motif prefers disorder to order upon phosphorylation of Ser 552 675 and Tyr670. In addition, the crucial secondary structural transition from α-helix to coil induced by phospho Ser552 and phospho Tyr670 of ß-Catenin ARM domain connecting helix C modifies conformational diversity and binding affinities of the complex interaction in functional regulation significantly. Moreover, the post phosphorylation disrupted the hydrogen bond interactions (Ser552-Arg549, Arg550-Asp546 and Ser675-Lys672) and abolished the residual alliance with hydrophobic interactions (Tyr670-Leu674) that easily interrupt in secondary structure packing as well as folding conformations connecting ARM and helix C (R10, 12 & R1C) compared to unphosphorylation. Our integrated computational analysis may help in shedding light on understanding the induced folding and unfolding pattern due to motif phosphorylations. Overall, our results provide an atomistic structural description of the way phosphorylation facilitates conformational and dynamic changes in ß-Catenin, a fundamental molecular switch mechanism in triple-negative breast cancer pathogenesis.


Asunto(s)
Procesamiento Proteico-Postraduccional , beta Catenina/metabolismo , Humanos , Simulación de Dinámica Molecular , Fosforilación , Conformación Proteica , Dominios Proteicos , Serina/química , Tirosina/química , beta Catenina/química
19.
Phytomedicine ; 81: 153428, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33341025

RESUMEN

BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is a fatal and progressive fibrotic lung disease lacking a validated and effective therapy. Aberrant activation of the Wnt/ß-catenin signaling cascade plays the key role in the pathogenesis of IPF. Betulinic acid is a natural pentacyclic triterpenoid molecule that has excellent antitumor and antiviral activities. HYPOTHESIS: We hypothesized that BA has an anti-pulmonary fibrosis effect mediated by the suppression of the Wnt/ß-catenin pathway. Study design Pulmonary fibrosis markers were detected in vitro and in vivo to confirm the antifibrotic effect of BA. The Wnt/ß-catenin pathway-related proteins were overexpressed to determine the effect of BA on Wnt signaling. METHODS AND RESULTS: BA dose-dependently inhibited Wnt3a-induced fibroblast activation in vitro. Moreover, BA decreased Wnt3a- and LiCl-induced transcriptional activity, as assessed by the TOPFlash assay in fibroblasts, and repressed the expression of the Wnt target genes cyclin D1, axin 2, and S100A4. Further investigation indicated that BA restrained the nuclear accumulation of ß-catenin, mainly by increasing the phospho-ß-catenin ratio (S33/S37/T41 and S45), inhibited the phosphorylation of DVL2 and LRP, and decreased the levels of Wnt3a and LRP6. In agreement with the results of the in vitro assays, the in vivo experiments indicated that BA significantly decreased bleomycin-induced pulmonary fibrosis in mice and suppressed myofibroblast activation by inhibiting Wnt/ß-catenin signaling. CONCLUSION: BA may directly interfere with the Wnt/ß-catenin pathway to subsequently repress myofibroblast activation and pulmonary fibrosis.


Asunto(s)
Fibrosis Pulmonar Idiopática/tratamiento farmacológico , Triterpenos Pentacíclicos/farmacología , Proteínas Wnt/metabolismo , beta Catenina/metabolismo , Animales , Bleomicina/toxicidad , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Fibrosis Pulmonar Idiopática/inducido químicamente , Fibrosis Pulmonar Idiopática/metabolismo , Fibrosis Pulmonar Idiopática/patología , Masculino , Ratones Endogámicos C57BL , Miofibroblastos/efectos de los fármacos , Miofibroblastos/metabolismo , Miofibroblastos/patología , Fosforilación/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Vía de Señalización Wnt/efectos de los fármacos
20.
Phytomedicine ; 80: 153373, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33096451

RESUMEN

BACKGROUND: Hepatocellular Carcinoma (HCC) is extremely aggressive and presents low rates of response to the available chemotherapeutic agents. Many studies have focused on the search for alternative low-cost natural compounds with antiproliferative potential that selectively respond to liver cancer cells. PURPOSE: This study assessed the in vitro direct action of trans-chalcone (TC) on cells of the human HCC HuH7.5 cell line. METHODS: We subjected the HuH7.5 tumor cells to TC treatment at increasing concentrations (12.5-100 µM) for 24 and 48 h. Cell viability was verified through MTT and 50% inhibitory concentration of cells (IC50 23.66 µM) was determined within 48 h. We quantified trypan blue proliferation and light microscopy, ROS production, mitochondrial depolarization and autophagy, cell cycle analysis, and apoptosis using Muse® cell analyzer and immunocytochemical markings of p53 and ß-catenin. RESULTS: Data showed an effective dose- and time-dependent TC-cytotoxic action at low micromolar concentrations without causing toxicity to non-cancerous cells, such as erythrocytes. TC-treatment caused mitochondrial membrane damage and cell cycle G0/G1 phase arrest, increasing the presence of the p53 protein and decreasing ß-catenin, in addition, to inducing cell death by autophagy. Additionally, TC decreased the metastatic capacity of HuH7.5, which affected the migration/invasion of this type of cell. CONCLUSION: In vitro TC activity in the human HCC HuH7.5 tumor cell line is shown to be a potential molecule to develop new therapies to repair the p53 pathway and prevent the overexpression of Wnt/ß-catenin tumor development inducing autophagy cell death and decreasing metastatic capacity of HuH7.5 cell line.


Asunto(s)
Carcinoma Hepatocelular/tratamiento farmacológico , Chalcona/farmacología , Chalconas/farmacología , Neoplasias Hepáticas/tratamiento farmacológico , Proteína p53 Supresora de Tumor/metabolismo , beta Catenina/metabolismo , Apoptosis/efectos de los fármacos , Autofagia/efectos de los fármacos , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patología , Puntos de Control del Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Regulación hacia Abajo/efectos de los fármacos , Humanos , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patología , Regulación hacia Arriba/efectos de los fármacos
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