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1.
Nat Commun ; 11(1): 4951, 2020 10 02.
Artículo en Inglés | MEDLINE | ID: mdl-33009382

RESUMEN

Immunogenic cell death (ICD) and tumour-infiltrating T lymphocytes are severely weakened by elevated reactive oxygen species (ROS) in the tumour microenvironment. It is therefore of critical importance to modulate the level of extracellular ROS for the reversal of immunosuppressive environment. Here, we present a tumour extracellular matrix (ECM) targeting ROS nanoscavenger masked by pH sensitive covalently crosslinked polyethylene glycol. The nanoscavenger anchors on the ECM to sweep away the ROS from tumour microenvironment to relieve the immunosuppressive ICD elicited by specific chemotherapy and prolong the survival of T cells for personalized cancer immunotherapy. In a breast cancer model, elimination of the ROS in tumour microenvironment elicited antitumour immunity and increased infiltration of T lymphocytes, resulting in highly potent antitumour effect. The study highlights a strategy to enhance the efficacy of cancer immunotherapy by scavenging extracellular ROS using advanced nanomaterials.


Asunto(s)
Antineoplásicos/farmacología , Espacio Extracelular/metabolismo , Depuradores de Radicales Libres/metabolismo , Muerte Celular Inmunogénica , Especies Reactivas de Oxígeno/metabolismo , Animales , Línea Celular Tumoral , Células Dendríticas/efectos de los fármacos , Células Dendríticas/metabolismo , Proteína HMGB1/metabolismo , Muerte Celular Inmunogénica/efectos de los fármacos , Ratones Endogámicos BALB C , Tamaño de la Partícula , Polietilenglicoles/química , Microambiente Tumoral/efectos de los fármacos
2.
Nat Commun ; 11(1): 4935, 2020 10 01.
Artículo en Inglés | MEDLINE | ID: mdl-33004797

RESUMEN

Gramicidin A (1) is a peptide antibiotic that disrupts the transmembrane ion concentration gradient by forming an ion channel in a lipid bilayer. Although long used clinically, it is limited to topical application because of its strong hemolytic activity and mammalian cytotoxicity, likely arising from the common ion transport mechanism. Here we report an integrated high-throughput strategy for discovering analogues of 1 with altered biological activity profiles. The 4096 analogue structures are designed to maintain the charge-neutral, hydrophobic, and channel forming properties of 1. Synthesis of the analogues, tandem mass spectrometry sequencing, and 3 microscale screenings enable us to identify 10 representative analogues. Re-synthesis and detailed functional evaluations find that all 10 analogues share a similar ion channel function, but have different cytotoxic, hemolytic, and antibacterial activities. Our large-scale structure-activity relationship studies reveal the feasibility of developing analogues of 1 that selectively induce toxicity toward target organisms.


Asunto(s)
Antibacterianos/farmacología , Descubrimiento de Drogas/métodos , Gramicidina/análogos & derivados , Ensayos Analíticos de Alto Rendimiento/métodos , Animales , Antibacterianos/química , Línea Celular Tumoral , Química Farmacéutica , Eritrocitos , Estudios de Factibilidad , Bacterias Grampositivas/efectos de los fármacos , Gramicidina/química , Gramicidina/farmacología , Hemólisis/efectos de los fármacos , Concentración 50 Inhibidora , Ratones , Pruebas de Sensibilidad Microbiana , Estructura Molecular , Conejos , Relación Estructura-Actividad , Espectrometría de Masas en Tándem
3.
Nat Commun ; 11(1): 5077, 2020 10 08.
Artículo en Inglés | MEDLINE | ID: mdl-33033240

RESUMEN

Although substantial progress has been made in cancer biology and treatment, clinical outcomes of bladder carcinoma (BC) patients are still not satisfactory. The tumor microenvironment (TME) is a potential target. Here, by single-cell RNA sequencing on 8 BC tumor samples and 3 para tumor samples, we identify 19 different cell types in the BC microenvironment, indicating high intra-tumoral heterogeneity. We find that tumor cells down regulated MHC-II molecules, suggesting that the downregulated immunogenicity of cancer cells may contribute to the formation of an immunosuppressive microenvironment. We also find that monocytes undergo M2 polarization in the tumor region and differentiate. Furthermore, the LAMP3 + DC subgroup may be able to recruit regulatory T cells, potentially taking part in the formation of an immunosuppressive TME. Through correlation analysis using public datasets containing over 3000 BC samples, we identify a role for inflammatory cancer-associated fibroblasts (iCAFs) in tumor progression, which is significantly related to poor prognosis. Additionally, we characterize a regulatory network depending on iCAFs. These results could help elucidate the protumor mechanisms of iCAFs. Our results provide deep insight into cancer immunology and provide an essential resource for drug discovery in the future.


Asunto(s)
Fibroblastos/patología , Inflamación/patología , Análisis de Secuencia de ARN , Análisis de la Célula Individual , Vejiga Urinaria/patología , Área Bajo la Curva , Fibroblastos Asociados al Cáncer/metabolismo , Fibroblastos Asociados al Cáncer/patología , Línea Celular Tumoral , Polaridad Celular , Proliferación Celular , Citocinas/metabolismo , Variaciones en el Número de Copia de ADN/genética , Células Dendríticas/metabolismo , Redes Reguladoras de Genes , Humanos , Ligandos , Glicoproteínas de la Membrana Asociadas a los Lisosomas/metabolismo , Monocitos/patología , Células Mieloides/patología , Proteínas de Neoplasias/metabolismo , Linfocitos T Reguladores/inmunología , Microambiente Tumoral , Vejiga Urinaria/inmunología
4.
Nat Commun ; 11(1): 5060, 2020 10 08.
Artículo en Inglés | MEDLINE | ID: mdl-33033246

RESUMEN

Fusion oncogenes (FOs) are common in many cancer types and are powerful drivers of tumor development. Because their expression is exclusive to cancer cells and their elimination induces cell apoptosis in FO-driven cancers, FOs are attractive therapeutic targets. However, specifically targeting the resulting chimeric products is challenging. Based on CRISPR/Cas9 technology, here we devise a simple, efficient and non-patient-specific gene-editing strategy through targeting of two introns of the genes involved in the rearrangement, allowing for robust disruption of the FO specifically in cancer cells. As a proof-of-concept of its potential, we demonstrate the efficacy of intron-based targeting of transcription factors or tyrosine kinase FOs in reducing tumor burden/mortality in in vivo models. The FO targeting approach presented here might open new horizons for the selective elimination of cancer cells.


Asunto(s)
Sistemas CRISPR-Cas/genética , Neoplasias/genética , Fusión de Oncogenes/genética , Animales , Secuencia de Bases , Línea Celular Tumoral , Proliferación Celular/genética , Doxorrubicina/uso terapéutico , Proteínas de Fusión bcr-abl/genética , Eliminación de Gen , Sitios Genéticos , Inestabilidad Genómica , Células HEK293 , Humanos , Intrones/genética , Ratones Desnudos , Neoplasias/tratamiento farmacológico , Neoplasias/patología , Proteínas de Fusión Oncogénica/genética , ARN Guia/metabolismo , Reproducibilidad de los Resultados , Ensayos Antitumor por Modelo de Xenoinjerto
5.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 28(5): 1451-1458, 2020 Oct.
Artículo en Chino | MEDLINE | ID: mdl-33067936

RESUMEN

OBJECTIVE: To investigate the proliferation inhibition and pro-apoptotic effect of Huaier aqueous extract combined with routine chemotherapeutic drugs including Vincristine (VCR), Daunorubicin (DNR), L-aspartase (L-Asp) on human acute lymphoblastic leukemia cell lines Nalm-6 and Sup-B15. METHODS: Nalm-6 and Sup-B15 cell lines were treated with different concentrations of Huaier aqueous extract and chemotherapeutics including VCR, DNR, L-Asp alone or in combination for 48 h, and the growth inhibitory effect and IC50 values (the half maximal inhibitory concentration) were detected by CCK-8. Jin's formula was used to estimated the synergistic effect of these combinations. Apoptosis rates of Nalm-6 and Sup-B15 cells and expression of apoptosis-related proteins BAX, BCL-2, cleaved Caspase-3 were determined by flow cytometry and Western blot respectivcly. RESULTS: Huaier aqueous extract, VCR, DNR and L-Asp had inhibition effect on Nalm-6 and Sup-B15 cell lines. The inhibition rate of Huaier aqueous extract combined with VCR, DNR and L-Asp were all higher than those of each dug alone (P<0.05) and the combination index (q) was between 0.85 and 1.15 or greater than 1.15. The two kinds of drugs showed had additive or synergistic effects. The results of flow cytometry showed that the cell apoptosis rates in combined treatment group were higher than those of each drug alone (P<0.05). The results of Western blot revealed that Huaier aqueous extract and VCR all decreased protein expression of BCL-2 (P<0.05) and increase protein expression of BAX (P<0.05) and cleaved Caspase-3 (P<0.05) in Nalm-6 and Sup-B15 cells. Compared with Huaier aqueous extract or VCR alone, the effect of two drug combination were more significant. DNR down-regulated protein expression of BCL-2 (P<0.05) and up-regulated cleaved Caspase-3 (P<0.05). However, it had no effect on the expression of BAX in Nalm-6 and Sup-B15 cells. When it was combined with Huaier aqueous extract, the expression of cleaved Caspase-3 and BCL-2 showed more significant changes. The expression of BAX in combined treated group did not show significant difference, compared with group treated with Huaier aqueous extract in Nalm-6 and Sup-B15 cells. L-Asp did not show significant effect on the three apoptosis-related proteins and there was no significant difference between the combination group and the Huaier aqueous extract group. CONCLUSION: the combination of Huaier aqueous extract and VCR, DNR, L-Asp shows additive or synergistic effects on human acute lymphoblastic leukemia cell lines Nalm-6 and Sup-B15.


Asunto(s)
Antineoplásicos , Leucemia-Linfoma Linfoblástico de Células Precursoras , Antineoplásicos/uso terapéutico , Línea Celular Tumoral , Mezclas Complejas/uso terapéutico , Humanos , Leucemia-Linfoma Linfoblástico de Células Precursoras/tratamiento farmacológico , Trametes
6.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 28(5): 1558-1562, 2020 Oct.
Artículo en Chino | MEDLINE | ID: mdl-33067954

RESUMEN

AbstractObjective: To investigate the in vitro biological characteristics of side-population(SP) cells in mantle cell lymphoma(MCL). METHODS: The SP cells in JeKo-1 cells were enriched by continuous culture and sorted by flow cytometry(FCM), and the expression of CD19 and IgM were analyzed. The differences of gene expression and their self-renewal ability between SP cells and non-SP cells were estimated by qRT-PCR and colony-forming assay(CFA) respectively. RESULTS: SP cells in JeKo-1 cells could be enriched by continuous sorting and in vitro cultureing, and it was found that CD19-/IgM- accounted for (2.79±0.82)%,CD19-/IgM+accounted for (70.99±13.61)%,CD19+/IgM-accounted for (0.55±0.25)%, and CD19+/IgM+accounted for (25.67±14)%. Bmi-1,CD133, C-MYC and Nanog genes were highly expressed in SP cells, but were low expressed in non-SP cells (P<0.05). Compared with non-SP cells, SP cells had stronger colony forming ability (P<0.05). CONCLUSION: JeKo-1 cell line of mantle cell lymphoma contains SP cells, which can be enriched by continuous sorting and culture for research. The SP cells in JeKo-1 contain four different cell populations with different immunophenotypes: CD19-/IgM-、CD19-/IgM+、CD19+/IgM-、CD19+/IgM+, and show stronger gene expression of stem cells and self-renewal ability in vitro.


Asunto(s)
Células Madre Neoplásicas , Células de Población Lateral , Línea Celular Tumoral , Citometría de Flujo , Inmunofenotipificación
7.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 28(5): 1578-1584, 2020 Oct.
Artículo en Chino | MEDLINE | ID: mdl-33067957

RESUMEN

OBJECTIVE: To investigate the potential mechanisms of miR-224 affecting the proliferation and invasion of diffuse large B-cell lymphoma (DLBCL) cells. METHODS: The blood samples of 76 DLBCL patients(DLBCL group) diagnosed at the First Affiliated Hospital of Kunming Medical University from June 2011 to December 2017, and 41 healthy persons of physical examination (normal control group) as well as human lymphatic endothelial cells (HELC) and DLBCL cell lines HBL1, OCI-LY10, OCI-LY8, OCI-LY19 were collected. The expression of miR-224 and PIK3CD mRNA was measured by RT-qPCR. The proliferation of HBL1 cells was detected by CCK-8 method and colony formation test. The invasion ability of HBL1 cells was detected by Transwell test. Dual-luciferase reporter gene assay was used to verify the relationship between miR-224 and PIK3CD. The expression of PIK3CD protein was measured by Western blot. RESULTS: The expression of miR-224 in blood of DLBCL patients was very significantly lower than that in normal control group (P<0.01). The overexpression of miR-224 significantly decreased proliferation and invasion of HBL1 cells (all P<0.01). PIK3CD was a target gene of miR-224. Knockdown of PIK3CD significantly suppressed the proliferation and invasion of HBL1 cells (all P<0.01). CONCLUSION: MiR-224 plays a key role in the progression of DLBCL, and the proliferation and invasion of HBL1 cells can be suppressed by targeted inhibition of PIK3CD.


Asunto(s)
Linfoma de Células B Grandes Difuso , MicroARNs , Línea Celular Tumoral , Proliferación Celular , Fosfatidilinositol 3-Quinasa Clase I/genética , Células Endoteliales , Humanos , Linfoma de Células B Grandes Difuso/genética , MicroARNs/genética
8.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 28(5): 1605-1610, 2020 Oct.
Artículo en Chino | MEDLINE | ID: mdl-33067961

RESUMEN

OBJECTIVE: To investigate the effect and possible mechanism of up-regulation of p-Akt by doxycycline (DOX) on myeloma cell line H929. METHODS: Multiple myeloma cell line H929 was treated with DOX at different concentrations for different times, and cell proliferation rate was measured by CCK-8 assay. The protein expression level of p-Akt, PTEN, p-PDK1, p-mTOR, p-GSK-3ß, and p-BAD was analyzed by Western blot. The mRNA levels of mTOR, BCL-2, and NF-κB was analyzed by RT-PCR. PI3K inhibitor Wortmannin was used to antagonize the up-regulation of p-Akt, and the cell proliferation and p-Akt protein expression level were analyzed by CCK-8 assay and Western blot respectively. RESULTS: DOX could inhibit the proliferation of H929 cells and up-regulate the expression of p-Akt at the same time. The protein levels of both p-PDK1 and PTEN in H929 cells did not alter significantly during DOX treatment. The expressions of p-BAD and p-GSK-3ß were up-regulated in H929 cells after treated with DOX, but the expression of p-mTOR was not altered. The mRNA levels of mTOR, BCL-2, and NF-κB in H929 were all down-regulated in H929 cells during DOX treatment. The effect up-regulating p-Akt level by DOX was suppressed when DOX combined with PI3K inhibitor Wortmannin and Wortmannin could enhance the inhibitory effect of DOX in H929 cells. CONCLUSION: DOX can activate PI3K/Akt signaling pathway in H929 cells, and antagonizing this effect of DOX may enhance its cytotoxicity to myeloma cells.


Asunto(s)
Mieloma Múltiple , Apoptosis , Línea Celular Tumoral , Doxiciclina/farmacología , Glucógeno Sintasa Quinasa 3 beta , Humanos , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de Señal , Regulación hacia Arriba
9.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 28(5): 1654-1660, 2020 Oct.
Artículo en Chino | MEDLINE | ID: mdl-33067969

RESUMEN

OBJECTIVE: To investigate the effects of metformin on the proliferation of AML-MDS cells (SKM-1 cells) and its related mechanisms. METHODS: CCK-8 was used to test the cell proliferation; Flow cytometry was used to detect the cell apoptosis and cell cycle; Western blot was used to test the expression level of AMPK and cell cycle regulatory proteins. RESULTS: Metformin could inhibit the proliferation of SKM-1 cells, which may be attributed to metformin-induced cell cycle arrest in G0/G1 but not to metformin induced cell apoptosis. The expression levels of G1-related protein CyclinD1 and CDK4 were down-regulated, while the expression levels of P53, P21CIP1 and P27kIP1 were up-regulated. Moreover, the phosphorylation level of AMPK was up-regulated. CONCLUSION: Metformin inhibits the proliferation of SKM-1 cells, which may relate with AMPK-induced cell cycle arrest. However, future studies are necessary to further explore the related mechanisms.


Asunto(s)
Metformina , Apoptosis , Puntos de Control del Ciclo Celular , Línea Celular Tumoral , Proliferación Celular , Metformina/farmacología
10.
Wei Sheng Yan Jiu ; 49(5): 780-784, 2020 Sep.
Artículo en Chino | MEDLINE | ID: mdl-33070824

RESUMEN

OBJECTIVE: Investigate the estrogen receptor expression in human thyroid squamous cell carcinoma SW579 and the effects of genistein on the apoptosis and cycle of SW579 and its mechnism. METHODS: The real-time PCR was applied to detect the expression of estrogen receptor(ER)α、ERß and G protein-coupled receptor(GPR)30 in human thyroid squamous cell line SW579; MTT was used to test the effect of genistein on cell proliferation in the SW579 cells before and after blocking GPR30; flow cytometry was explorited to measure the effect of genistein on the cell cycle and apoptosis in the SW579 was detected before and after blocking GPR30. RESULTS: The high concentration of genistein promoted the expression of ERß and GPR30 in the SW579 cells, but ERα was not expressed. The specific blocking of GPR30, the cell proliferation was aboviously inhibited by genistein in the SW579 cells and the cell apoptosis was markedly promoted after the GPR30 was blocked; The cell cycle was mainly blocked in G_2/M phase. CONCLUSION: Genistein can obiviously promote the cell proliferation in the SW579 cells, which may be related to the action of GPR30.


Asunto(s)
Genisteína , Neoplasias de la Tiroides , Apoptosis , Ciclo Celular , División Celular , Línea Celular Tumoral , Células Epiteliales , Genisteína/farmacología , Humanos
11.
Nat Commun ; 11(1): 4980, 2020 10 05.
Artículo en Inglés | MEDLINE | ID: mdl-33020477

RESUMEN

The functions of the proto-oncoprotein c-Myc and the tumor suppressor p53 in controlling cell survival and proliferation are inextricably linked as "Yin and Yang" partners in normal cells to maintain tissue homeostasis: c-Myc induces the expression of ARF tumor suppressor (p14ARF in human and p19ARF in mouse) that binds to and inhibits mouse double minute 2 homolog (MDM2) leading to p53 activation, whereas p53 suppresses c-Myc through a combination of mechanisms involving transcriptional inactivation and microRNA-mediated repression. Nonetheless, the regulatory interactions between c-Myc and p53 are not retained by cancer cells as is evident from the often-imbalanced expression of c-Myc over wildtype p53. Although p53 repression in cancer cells is frequently associated with the loss of ARF, we disclose here an alternate mechanism whereby c-Myc inactivates p53 through the actions of the c-Myc-Inducible Long noncoding RNA Inactivating P53 (MILIP). MILIP functions to promote p53 polyubiquitination and turnover by reducing p53 SUMOylation through suppressing tripartite-motif family-like 2 (TRIML2). MILIP upregulation is observed amongst diverse cancer types and is shown to support cell survival, division and tumourigenicity. Thus our results uncover an inhibitory axis targeting p53 through a pan-cancer expressed RNA accomplice that links c-Myc to suppression of p53.


Asunto(s)
Neoplasias/patología , Proteínas Proto-Oncogénicas c-myc/metabolismo , ARN Largo no Codificante/metabolismo , Proteína p53 Supresora de Tumor/metabolismo , Carcinogénesis , Proteínas Portadoras/genética , Proteínas Portadoras/metabolismo , Línea Celular Tumoral , Supervivencia Celular , Regulación Neoplásica de la Expresión Génica , Humanos , Neoplasias/genética , Neoplasias/metabolismo , Regiones Promotoras Genéticas , Unión Proteica , Proteínas Proto-Oncogénicas c-myc/genética , ARN Largo no Codificante/genética , Sumoilación , Proteína p53 Supresora de Tumor/genética , Ubiquitinación
12.
Nat Commun ; 11(1): 4983, 2020 10 05.
Artículo en Inglés | MEDLINE | ID: mdl-33020492

RESUMEN

The energetic demands of a cell are believed to increase during mitosis, but the rates of ATP synthesis and consumption during mitosis have not been quantified. Here, we monitor mitochondrial membrane potential of single lymphocytic leukemia cells and demonstrate that mitochondria hyperpolarize from the G2/M transition until the metaphase-anaphase transition. This hyperpolarization was dependent on cyclin-dependent kinase 1 (CDK1) activity. By using an electrical circuit model of mitochondria, we quantify mitochondrial ATP synthesis rates in mitosis from the single-cell time-dynamics of mitochondrial membrane potential. We find that mitochondrial ATP synthesis decreases by approximately 50% during early mitosis and increases back to G2 levels during cytokinesis. Consistently, ATP levels and ATP synthesis are lower in mitosis than in G2 in synchronized cell populations. Overall, our results provide insights into mitotic bioenergetics and suggest that cell division is not a highly energy demanding process.


Asunto(s)
Adenosina Trifosfato/biosíntesis , División Celular , Metabolismo Energético , Animales , Proteína Quinasa CDC2/metabolismo , Línea Celular Tumoral , Citocinesis , Ratones , Mitocondrias/metabolismo , Mitosis , Modelos Biológicos
13.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 28(5): 1598-1604, 2020 Oct.
Artículo en Chino | MEDLINE | ID: mdl-33067960

RESUMEN

OBJECTIVE: To detect the expression of MutT homolog 1 (MTH1) in CD138-negative cells (CD138-) and CD138-positive cells (CD138+) cells of the patients with multiple myeloma (MM), and to explore the effect of MTH1 inhibitor TH588 on cell morphology, proliferation and apoptosis of MM cell U266. METHODS: CD138- and CD138+ cells of MM patients were isolated, and RNA was extracted. The expression of NUDT family was detected by Q-PCR. MM cell line U266 was used to observe the effect of IL-6 on MTH1 expression. Using fluorescence microscopy to observe the morphological changes of U266 after treatment by TH588 for 48 hours. DAPI staining was used to investigate the nuclear change. Luciferase was used to detect the effect of TH588 on U266 cell proliferation. After treatment with TH588 for 48 h, the change of apoptosis in MM U266 cells was detected by flow cytometer. RESULTS: In some MM patients, the expression of MTH1, NUDT2, NUDT5 and NUDT21 in CD138+ cells was higher than that in CD138- (P<0.05). The luciferase report showed that after treatment of U266 cells with TH588, the fluorescence intensity was significantly lower than that of the control group, and the fluorescence intensity decreased with the increase of drug concentration (r=-0.91). IL-6 could increase the expression of MTH1. Fluorescence microscopy showed that after TH588 treatment of U266 cells for 48 hours, the cells appeared shrinking, smaller, and irregular in shape. After DAPI staining, the TH588-treated cells showed apoptosis characteristics, such as nuclear shrinkage, uneven staining, petal, and radial shape. Flow cytometry showed that the proportion of U266 viable cells decreased significantly after treatment with TH588 for 48 h (P<0.05). CONCLUSION: MTH1 highly expresses in CD138+ cells of some MM patients. MTH1 expression can increase in U266 cells treated by IL-6. The MTH1 inhibitor TH588 possesses proliferation-inhibitory effect and apoptosis-inducing effect on MM cell U266.


Asunto(s)
Mieloma Múltiple , Apoptosis , Línea Celular Tumoral , Proliferación Celular , Factor de Especificidad de Desdoblamiento y Poliadenilación , Enzimas Reparadoras del ADN , Citometría de Flujo , Humanos , Monoéster Fosfórico Hidrolasas
14.
Nat Commun ; 11(1): 4912, 2020 09 30.
Artículo en Inglés | MEDLINE | ID: mdl-32999275

RESUMEN

Most signals detected by genome-wide association studies map to non-coding sequence and their tissue-specific effects influence transcriptional regulation. However, key tissues and cell-types required for functional inference are absent from large-scale resources. Here we explore the relationship between genetic variants influencing predisposition to type 2 diabetes (T2D) and related glycemic traits, and human pancreatic islet transcription using data from 420 donors. We find: (a) 7741 cis-eQTLs in islets with a replication rate across 44 GTEx tissues between 40% and 73%; (b) marked overlap between islet cis-eQTL signals and active regulatory sequences in islets, with reduced eQTL effect size observed in the stretch enhancers most strongly implicated in GWAS signal location; (c) enrichment of islet cis-eQTL signals with T2D risk variants identified in genome-wide association studies; and (d) colocalization between 47 islet cis-eQTLs and variants influencing T2D or glycemic traits, including DGKB and TCF7L2. Our findings illustrate the advantages of performing functional and regulatory studies in disease relevant tissues.


Asunto(s)
Glucemia/genética , Diabetes Mellitus Tipo 2/genética , Predisposición Genética a la Enfermedad , Islotes Pancreáticos/metabolismo , Sitios de Carácter Cuantitativo , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Animales , Glucemia/metabolismo , Línea Celular Tumoral , Estudios de Cohortes , Diabetes Mellitus Tipo 2/sangre , Diacilglicerol Quinasa/genética , Diacilglicerol Quinasa/metabolismo , Elementos de Facilitación Genéticos , Femenino , Regulación de la Expresión Génica , Estudio de Asociación del Genoma Completo , Humanos , Masculino , Ratones , Persona de Mediana Edad , Polimorfismo de Nucleótido Simple , RNA-Seq , Análisis de Secuencia de ADN , Proteína 2 Similar al Factor de Transcripción 7/genética , Proteína 2 Similar al Factor de Transcripción 7/metabolismo , Adulto Joven
15.
Nat Commun ; 11(1): 5079, 2020 10 08.
Artículo en Inglés | MEDLINE | ID: mdl-33033234

RESUMEN

Tumor heterogeneity and lack of knowledge about resistant cell states remain a barrier to targeted cancer therapies. Basal cell carcinomas (BCCs) depend on Hedgehog (Hh)/Gli signaling, but can develop mechanisms of Smoothened (SMO) inhibitor resistance. We previously identified a nuclear myocardin-related transcription factor (nMRTF) resistance pathway that amplifies noncanonical Gli1 activity, but characteristics and drivers of the nMRTF cell state remain unknown. Here, we use single cell RNA-sequencing of patient tumors to identify three prognostic surface markers (LYPD3, TACSTD2, and LY6D) which correlate with nMRTF and resistance to SMO inhibitors. The nMRTF cell state resembles transit-amplifying cells of the hair follicle matrix, with AP-1 and TGFß cooperativity driving nMRTF activation. JNK/AP-1 signaling commissions chromatin accessibility and Smad3 DNA binding leading to a transcriptional program of RhoGEFs that facilitate nMRTF activity. Importantly, small molecule AP-1 inhibitors selectively target LYPD3+/TACSTD2+/LY6D+ nMRTF human BCCs ex vivo, opening an avenue for improving combinatorial therapies.


Asunto(s)
Carcinoma Basocelular/metabolismo , Proteínas Hedgehog/metabolismo , Transducción de Señal , Neoplasias Cutáneas/metabolismo , Factor de Transcripción AP-1/metabolismo , Factor de Crecimiento Transformador beta/metabolismo , Animales , Línea Celular Tumoral , Núcleo Celular/metabolismo , Cromatina/metabolismo , ADN de Neoplasias/metabolismo , Resistencia a Antineoplásicos , Matriz Extracelular/metabolismo , Ontología de Genes , Factores de Intercambio de Guanina Nucleótido/metabolismo , Folículo Piloso/metabolismo , Humanos , Ratones , Ratones Endogámicos C57BL , Células 3T3 NIH , Proteínas de Neoplasias/metabolismo , Unión Proteica , Proteína smad3/metabolismo , Transactivadores/metabolismo , Regulación hacia Arriba
16.
Shanghai Kou Qiang Yi Xue ; 29(3): 267-274, 2020 Jun.
Artículo en Chino | MEDLINE | ID: mdl-33043343

RESUMEN

PURPOSE: To investigate the molecular mechanism of LncRNA NEAT1 regulating proliferation, migration and invasion of tongue squamous cell carcinoma cells by regulating miR-339-5p/ITGA3 axis. METHODS: qRT-PCR and Western blot were used to detect the expression of NEAT1, miR-339-5p, ITGA3 mRNA and ITGA3 protein in 25 cases of human tongue squamous cell carcinoma, its corresponding adjacent tissues, human normal oral mucosal cell line HOK and human tongue squamous cell carcinoma cell lines TSCCA, CAL27, SCC15 and HN13. CAL27 cell lines that inhibited NEAT1 and overexpressed miR-339-5p were constructed, respectively. Cell viability was detected by MTT assay, cell numbers of migration and invasion were detected by Transwell assay, and the expression of Cyclin D1 and MMP-9 proteins were detected by Western blotting. The dual luciferase reporter gene was used to verify the targeting relationship of NEAT1, miR-339-5p and ITGA3, and the regulatory relationship was detected by Western blotting and qRT-PCR. SPSS 17.0 software package was used for statistical analysis of the data. RESULTS: Compared with normal human oral mucosal cell line HOK, the expression of NEAT1 and ITGA3 was up-regulated, while the expression of miR-339-5p was down-regulated in human tongue squamous cell carcinoma cell lines. Inhibition of NEAT1 or over-expression of miR-339-5p significantly inhibited proliferation, migration and invasion of CAL27 cells, and significantly inhibited expression of Cyclin D1 and MMP-9 proteins. Dual luciferase reporter gene assay confirmed that NEAT1 directly interacted with miR-339-5p and suppressed its expression. miR-339-5p negatively regulated ITGA3 expression. Inhibition of NEAT1 reversed the inhibitory effect of the inhibition of miR-339-5p on proliferation, migration and invasion of CAL27 cells. CONCLUSIONS: LncRNA NEAT1 promotes proliferation, migration and invasion of tongue squamous cell carcinoma cells by down-regulating miR-339-5p/ITGA3 axis.


Asunto(s)
Carcinoma de Células Escamosas , MicroARNs , ARN Largo no Codificante/fisiología , Carcinoma de Células Escamosas/genética , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Regulación Neoplásica de la Expresión Génica , Humanos , Integrina alfa3 , MicroARNs/genética , ARN Largo no Codificante/genética
17.
Signal Transduct Target Ther ; 5(1): 221, 2020 10 06.
Artículo en Inglés | MEDLINE | ID: mdl-33024073
18.
Beijing Da Xue Xue Bao Yi Xue Ban ; 52(5): 902-906, 2020 Oct 18.
Artículo en Chino | MEDLINE | ID: mdl-33047727

RESUMEN

OBJECTIVE: To investigate the effects of salinomycin on the proliferation and apoptosis of oral squamous carcinoma cells and to further understand the mechanisms of these effects. METHODS: The human oral squamous carcinoma cell line CAL-27 was cultured in different concentrations of salinomycin and cisplatin. After co-culture with 0, 1, 2, 4, 8, 16 and 32 µmol/L salinomycin or 0, 1.25, 2.5, 5, 10, 20, 40 and 80 µmol/L cisplatin for 24 hours and 48 hours, the proliferation of oral squamous carcinoma cells were detected by cell counting kit-8(CCK-8) assay. After being exposed to 0, 2, 4, 8 µmol/L salinomycin and 0, 5, 10, 20 µmol/L cisplatin for 48 hours, the cell cycle of oral squamous carcinoma cells was detected by flow cytometry assay, and Western blot analysis was performed to analyze the expressions of cysteine-containing aspartate-specific proteases-3(Caspase-3), cysteine-containing aspartate-specific proteases-9(Caspase-9), poly ADP-ribose polymerase (PARP), protein kinase B (Akt) and phosphorylated protein kinase B (p-Akt) protein in oral squamous carcinoma cells. RESULTS: Both salinomycin and cisplatin significantly inhibited the proliferation of oral squamous cell carcinoma CAL-27 cells in a time- and dose-dependent manner. However, compared with the first-line chemotherapeutic drug cisplatin, salinomycin showed stronger anti-proliferation activity in oral squamous carcinoma cells than cisp-latin (P < 0.001). After being exposed to 8 µmol/L salinomycin, CAL-27 cells exhibited markedly higher proportion in quiescent/ first gap phases (40.40%±1.99% vs. 64.46%±0.90%, P < 0.05), and had a significantly lower proportion in synthesis phases and second gap / mitosis phases (24.32%±2.30% vs. 18.73%±0.61%, P < 0.05; 35.01%±1.24% vs. 16.54%±1.31%, P < 0.05) compared with the dimethyl sulfoxide control group; moreover cisplatin didn't show cell-cycle specific effect on CAL-27. Western blot proved that salinomycin could up-regulate the expressions of Caspase-3 and Caspase-9 protein in oral squamous cell carcinoma CAL-27 cells (P < 0.05). At the same time, the levels of PARP, Akt and p-Akt protein were down-regulated (P < 0.05). CONCLUSION: Compared with cisplatin, salinomycin has a better inhibitory effect on the proliferation of oral squamous carcinoma cells and blocks the cell cycle process at the quiescent / first gap phase. At the same time, salinomycin could trigger apoptosis of oral squamous carcinoma cells and the mechanism is associated with the Akt/p-Akt signaling pathway.


Asunto(s)
Antineoplásicos , Carcinoma de Células Escamosas , Neoplasias de la Boca , Antineoplásicos/farmacología , Antineoplásicos/uso terapéutico , Apoptosis/efectos de los fármacos , Carcinoma de Células Escamosas/tratamiento farmacológico , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Humanos , Neoplasias de la Boca/tratamiento farmacológico , Piranos
19.
Folia Biol (Praha) ; 66(3): 104-110, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-33069189

RESUMEN

Cancer development is a highly complicated process in which tumour growth depends on the development of its vascularization system. To support their own growth, tumour cells significantly modify their microenvironment. One of such modifications inflicted by tumours is stimulation of endothelial cell migration and proliferation. There is accumulating evidence that extracellular vesicles (EVs) secreted by tumour cells (tumour-derived EVs, TEVs) may be regarded as "messengers" with the potential for affecting the biological activities of target cells. Interaction of TEVs with different cell types occurs in an auto- and paracrine manner and may lead to changes in the function of the latter, e.g., promoting motility, proliferation, etc. This study analysed the proangiogenic activity of EVs derived from human pancreatic adenocarcinoma cell line (HPC-4, TEVHPC) in vitro and their effect in vivo on Matrigel matrix vascularization in severe combined immunodeficient (SCID) mice. TEVHPC enhanced proliferation of HPC-4 cells and induced their motility. Moreover, TEVHPC stimulated human umbilical vein endothelial cell (HUVEC) proliferation and migration in vitro. Additionally, TEVHPC influenced secretion of proangiogenic factors (IL-8, VEGF) by HUVEC cells and supported Matrigel matrix haemoglobinization in vivo. These data show that TEVs may support tumour propagation in an autocrine manner and may support vascularization of the tumour. The presented data are in line with the theory that tumour cells themselves are able to modulate the microenvironment via TEVs to maximize their growth potential.


Asunto(s)
Carcinoma Ductal Pancreático/patología , Vesículas Extracelulares/patología , Neoplasias Pancreáticas/patología , Animales , Comunicación Autocrina , División Celular , Línea Celular Tumoral , Quimiotaxis , Colágeno , Combinación de Medicamentos , Células Endoteliales de la Vena Umbilical Humana , Humanos , Interleucina-8/genética , Interleucina-8/metabolismo , Laminina , Ratones , Ratones SCID , Trasplante de Neoplasias , Neovascularización Patológica/etiología , Proteoglicanos , ARN Mensajero/biosíntesis , Microambiente Tumoral , Factor A de Crecimiento Endotelial Vascular/genética , Factor A de Crecimiento Endotelial Vascular/metabolismo
20.
Hua Xi Kou Qiang Yi Xue Za Zhi ; 38(5): 495-501, 2020 Oct 01.
Artículo en Chino | MEDLINE | ID: mdl-33085231

RESUMEN

OBJECTIVE: This study aims to study the effect of the enhancer of zeste homolog 2 (EZH2) inhibitor GSK126 on the proliferation and apoptosis of human tongue squamous cell carcinoma cells in vitro and explore its related mechanisms in order to obtain insights into the clinical treatment of tongue squamous cell carcinoma. METHODS: Different concentrations of GSK126 were applied to CAL-27 cells of tongue squamous cell carcinoma, and the effects of drugs on cell proliferation were detected through methyl thiazolyl tetrazolium (MTT) assay, colony formation assay, and 5-ethynyl-2'-deoxyuridine (EdU) fluorescence staining. Hoechst33342 fluorescence staining and the JC-1 method were used in observing apoptosis. The expression levels of extracellular regulated protein kinases (ERK), phospho-extracellular regulated protein kinases (p-ERK), Bax, Bcl-2, and Cleaved caspase-9 in Cal-27 cells were detected through Western blot. RESULTS: GSK126 inhibited CAL-27 cell proliferation and promoted apoptosis. GSK126 down-regulated the expression of p-ERK and Bcl-2 and increased the expression of Bax and Cleaved caspase-9 (P<0.05). CONCLUSIONS: GSK126 can inhibit the proliferation of CAL-27 cells in tongue squamous cell carcinoma and promote its apoptosis, and the related mechanism may be associated with the inhibition of the MEK/ERK signaling pathway and activation of the Bax/Bcl-2 pathway.


Asunto(s)
Apoptosis , Neoplasias de Cabeza y Cuello , Línea Celular Tumoral , Proliferación Celular , Proteína Potenciadora del Homólogo Zeste 2 , Humanos , Indoles , Piridonas
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