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1.
Chem Biol Interact ; 323: 109075, 2020 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-32229109

RESUMO

The use of orchids in herbal medicine has a very long history. Dendrobium species are known to produce a variety of secondary metabolites such as phenanthrens, bibenzyls, fluorenones and sesquiterpenes, and alkaloids and are responsible for their wide variety of medicinal properties. For decades, bibenzyls, which are the main bioactive components derived from Dendrobium species, have been subjected to extensive investigation as likely candidates for cancer treatment. The present study was undertaken to investigate the effect of moscatilin, a bibenzyl derivative from the orchid Dendrobium loddigesii on human melanoma cells. In A375 cells compound moscatilin showed a clear dose-response relationship in the range of 6.25-50 µM concentrations. In addition, we demonstrated an apoptotic response after treatment of cancer cells with this bibenzyl compound at 6.25 and 12.5 µM concentrations that probably involves PTEN activity, inhibition of Hsp70 expression and reactive oxygen species production. Alternatively, the inhibition of the caspase cascade at higher concentrations, 25 and 50 µM, correlated with additional reactive oxygen species increase, probably switched the mode of moscatilin-induced cell death from apoptosis to necrosis.


Assuntos
Apoptose/efeitos dos fármacos , Compostos de Benzil/uso terapêutico , Dendrobium/química , Melanoma/tratamento farmacológico , Melanoma/patologia , Compostos de Benzil/química , Compostos de Benzil/farmacologia , Caspase 3/metabolismo , Caspase 9/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , DNA de Neoplasias/metabolismo , Glutationa/metabolismo , Proteínas de Choque Térmico HSP70/metabolismo , Humanos , L-Lactato Desidrogenase/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Proteína X Associada a bcl-2/metabolismo
2.
Biol Res ; 53(1): 13, 2020 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-32293552

RESUMO

BACKGROUND: Gallbladder cancer (GBC) is the most common tumor of the biliary tract. The incidence of GBC shows a large geographic variability, being particularly frequent in Native American populations. In Chile, GBC represents the second cause of cancer-related death among women. We describe here the establishment of three novel cell lines derived from the ascitic fluid of a Chilean GBC patient, who presented 46% European, 36% Mapuche, 12% Aymara and 6% African ancestry. RESULTS: After immunocytochemical staining of the primary cell culture, we isolated and comprehensively characterized three independent clones (PUC-GBC1, PUC-GBC2 and PUC-GBC3) by short tandem repeat DNA profiling and RNA sequencing as well as karyotype, doubling time, chemosensitivity, in vitro migration capability and in vivo tumorigenicity assay. Primary culture cells showed high expression of CK7, CK19, CA 19-9, MUC1 and MUC16, and negative expression of mesothelial markers. The three isolated clones displayed an epithelial phenotype and an abnormal structure and number of chromosomes. RNA sequencing confirmed the increased expression of cytokeratin and mucin genes, and also of TP53 and ERBB2 with some differences among the three cells lines, and revealed a novel exonic mutation in NF1. The PUC-GBC3 clone was the most aggressive according to histopathological features and the tumorigenic capacity in NSG mice. CONCLUSIONS: The first cell lines established from a Chilean GBC patient represent a new model for studying GBC in patients of Native American descent.


Assuntos
Antígenos Glicosídicos Associados a Tumores/genética , Neoplasias da Vesícula Biliar/genética , Índios Sul-Americanos/genética , Animais , Antineoplásicos/farmacologia , Líquido Ascítico/metabolismo , Carcinogênese/genética , Testes de Carcinogenicidade , Linhagem Celular Tumoral/efeitos dos fármacos , Linhagem Celular Tumoral/metabolismo , Chile , Cisplatino/farmacologia , Células Clonais/efeitos dos fármacos , Células Clonais/metabolismo , Impressões Digitais de DNA , Desoxicitidina/análogos & derivados , Desoxicitidina/farmacologia , Células Epiteliais/metabolismo , Neoplasias da Vesícula Biliar/metabolismo , Perfilação da Expressão Gênica , Genes erbB-2/genética , Humanos , Queratina-19/genética , Queratina-7/genética , Masculino , Camundongos Endogâmicos NOD , Pessoa de Meia-Idade , Receptor ErbB-2/genética , Análise de Sequência de RNA , Células Tumorais Cultivadas , Proteína Supressora de Tumor p53/genética
3.
Gene ; 745: 144629, 2020 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-32229158

RESUMO

Small-cell lung cancer (SCLC) is the most invasive of all lung cancer subtypes, and is characterized by its rapid response to chemotherapy resistance. Overcoming chemotherapy resistance is therefore the key to treating SCLC. P53 is mutated in most SCLCs, which has an effect of enhancing chemotherapy resistance. Regulation of p53 proteins by a variety of post-translational modifications, such as acetylation, which affects their function. Acetylation and deacetylation of p53 may be potential targets for modulating chemosensitivity. Recent studies have shown that SIRT3 acts as a deacetylase that regulates acetylation of p53. However, whether SIRT3 can regulate the post-translational modification of mutant p53 has not been studied. In the present study, we found that SIRT3 can deacetylate mutant p53, thus reducing its expression, inducing apoptosis in SCLC cells, and increasing SCLC chemosensitivity. The relationship between SIRT3 and mutant p53 could be the basis of a new SCLC treatment approach.


Assuntos
Resistencia a Medicamentos Antineoplásicos/genética , Neoplasias Pulmonares/tratamento farmacológico , Sirtuína 3/metabolismo , Carcinoma de Pequenas Células do Pulmão/tratamento farmacológico , Proteína Supressora de Tumor p53/metabolismo , Acetilação , Animais , Apoptose/efeitos dos fármacos , Apoptose/genética , Linhagem Celular Tumoral , Cisplatino/farmacologia , Cisplatino/uso terapêutico , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Masculino , Camundongos , Mutação , Complexo de Endopeptidases do Proteassoma/metabolismo , Processamento de Proteína Pós-Traducional , Proteólise , Carcinoma de Pequenas Células do Pulmão/genética , Carcinoma de Pequenas Células do Pulmão/patologia , Ubiquitina/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto
4.
Toxicon ; 180: 11-17, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32259553

RESUMO

Ochratoxin A (OTA) is a toxic metabolite produced by Aspergillus and Penicillium fungus. OTA found in the human and animal tissues can contaminate many foods that we daily consume in our lives. It accumulates especially in kidney. Although OTA is known to cause cell cycle arrest, the molecular mechanisms underlying this effect have not been fully understood, yet. We aimed to investigate the molecular details of OTA induced inhibitory response in G1 - G1/S phase of cell cycle and also the regulatory role of p53 in OTA mediated cell cycle arrest in human proximal tubule epithelial cells, HK-2. For this purpose, Cyclin E1 and Cyclin D1 mRNA expressions and Cyclin D1, Cdk4 and Cdk2 protein expressions were evaluated in HK-2 cells transfected with either 50 nM control siRNA or p53 siRNA for 72 h in the absence or presence of OTA using RT-PCR and Western blot analyses, respectively. Our findings showed that mRNA expressions of Cyclin D1 and Cyclin E1 and protein expressions of Cyclin D1, Cdk4 and Cdk2 were inhibited in HK-2 cells treated with two different doses of OTA, 10 µM and 25 µM, for 24 h. However, the downregulation of p53 led to enhance OTA-mediated increase in mRNA expressions of Cyclin D1 and Cyclin E1 and protein expressions of Cyclin D1, Cdk4 and Cdk2 compared to control siRNA transfected HK-2 cells. Our findings strongly suggest that the cell cycle arresting effect of OTA also performs via a p53 mediated mechanism besides other possible mechanisms.


Assuntos
Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Ocratoxinas/toxicidade , Proteínas de Ciclo Celular/metabolismo , Linhagem Celular , Ciclina D1 , Ciclina E , Regulação para Baixo , Fase G1/efeitos dos fármacos , Humanos , Rim , Proteínas Oncogênicas , Fase S/efeitos dos fármacos , Testes de Toxicidade , Proteína Supressora de Tumor p53/metabolismo
5.
Emerg Microbes Infect ; 9(1): 761-770, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32228226

RESUMO

Circulating in China and 158 other countries and areas, the ongoing COVID-19 outbreak has caused devastating mortality and posed a great threat to public health. However, efforts to identify effectively supportive therapeutic drugs and treatments has been hampered by our limited understanding of host immune response for this fatal disease. To characterize the transcriptional signatures of host inflammatory response to SARS-CoV-2 (HCoV-19) infection, we carried out transcriptome sequencing of the RNAs isolated from the bronchoalveolar lavage fluid (BALF) and peripheral blood mononuclear cells (PBMC) specimens of COVID-19 patients. Our results reveal distinct host inflammatory cytokine profiles to SARS-CoV-2 infection in patients, and highlight the association between COVID-19 pathogenesis and excessive cytokine release such as CCL2/MCP-1, CXCL10/IP-10, CCL3/MIP-1A, and CCL4/MIP1B. Furthermore, SARS-CoV-2 induced activation of apoptosis and P53 signalling pathway in lymphocytes may be the cause of patients' lymphopenia. The transcriptome dataset of COVID-19 patients would be a valuable resource for clinical guidance on anti-inflammatory medication and understanding the molecular mechansims of host response.


Assuntos
Líquido da Lavagem Broncoalveolar , Quimiocinas/análise , Infecções por Coronavirus/genética , Citocinas/análise , Leucócitos Mononucleares , Pneumonia Viral/genética , Transcriptoma , Apoptose , Betacoronavirus , Infecções por Coronavirus/sangue , Infecções por Coronavirus/imunologia , Humanos , Linfopenia , Pandemias , Pneumonia Viral/sangue , Pneumonia Viral/imunologia , RNA-Seq , Transdução de Sinais , Proteína Supressora de Tumor p53
6.
Zhonghua Yi Xue Za Zhi ; 100(11): 823-827, 2020 Mar 24.
Artigo em Chinês | MEDLINE | ID: mdl-32234152

RESUMO

Objective: To observe the role of cerebrospinal fluid (CSF) TP53 gene mutation in lung cancer associated meningitis. Methods: A retrospective analysis was performed on 35 patients diagnosed with lung cancer associated meningitis at the Second Hospital of Hebei Medical University from December 2015 to December 2018.All patients underwent the next-generation sequencing of CSF, and TP53 gene was found to be mutant or wild type, including 23 patients with TP53 mutant type and 12 patients with TP53 wild type. The clinical characteristics, CSF leukocyte, protein, glucose, chloride, Karnofsky performance (KPS) and overall survival were observed. Results: Headache, nausea and vomiting were the main clinical manifestations in both groups.There were no significant differences in CSF pressure, leukocyte, biochemical indicators and KPS between the two groups. The average time from diagnosis of lung cancer to diagnosis of lung cancer associated meningitis in the TP53 mutant group was significantly shorter than that in the TP53 wild type group (5.79 months vs 25.5 months).The median survival time of patients in the TP53 mutant group from lung cancer diagnosis to the observation endpoint was 19.77 months, while it was 88.73 months in the TP53 wild type group, and the difference was statistically significant (P=0.043). Conclusions: Mutation in the tumor suppressor gene TP53 can be detected in the CSF of patients with lung cancer associated meningitis. Patients with such mutation have earlier meningeal metastasis and shorter median survival time.


Assuntos
Neoplasias Pulmonares , Meningite , Mutação , Proteína Supressora de Tumor p53/genética , Líquido Cefalorraquidiano , Genes p53 , Humanos , Neoplasias Pulmonares/complicações , Neoplasias Pulmonares/genética , Meningite/complicações , Meningite/genética , Metástase Neoplásica , Prognóstico , Estudos Retrospectivos , Análise de Sobrevida
7.
Asian Pac J Cancer Prev ; 21(3): 791-798, 2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-32212809

RESUMO

BACKGROUND: Cholangiocarcinoma (CCA), a common cancer in northeastern Thailand, is a severe disease with poor prognosis and short survival time following diagnosis. DNA damage in CCA is believed to be caused by liver fluke infection in combination with exposure to carcinogens. p53, a tumor suppressor, is the most mutated gene in human cancers including liver fluke-associated CCA. Hence, expression patterns of p53 and its target genes may be useful for diagnosis and/or prognosis of CCA patients. METHODS: Differential mRNA expression of p53 and its target genes, namely, FUCA1, ICAM2 MDM2, p21, PAI-1, S100A9, and WIP1 in CCA tissue samples (n = 30) relative to matched adjacent non-tumor tissues was determined by quantitative RT-PCR and compared to clinicopathological features. Level of p53 protein was determined by immunohistochemistry and correlated with the expression of its target genes. RESULTS: Immunohistochemistry showed elevation of p53 protein level in 77% of the cases, while RT-PCR showed downregulation of p53 mRNA and its seven target genes in 23% and 47-97% of the samples. PAI-1 was down-regulated in almost all CCA samples, thus highlighting it as a potential diagnostic marker for CCA. However, no significant clinical associations were found except for down-regulation of WIP1 that was significantly correlated with non-papillary type tissue (p-value = 0.001) and with high p53 protein level (p-value = 0.007). CONCLUSION: Our results demonstrated statistically significant association between down-regulation of WIP1 with non-papillary type and with high p53 protein level, and PAI-1 was down-regulated in almost all CCA. Therefore, expression level of WIP1 and PAI-1 may be useful for predicting p53 functional status and as a potential diagnostic marker of CCA, respectively.


Assuntos
Neoplasias dos Ductos Biliares/genética , Biomarcadores Tumorais/genética , Colangiocarcinoma/genética , Regulação Neoplásica da Expressão Gênica , Proteína Supressora de Tumor p53/genética , Idoso , Neoplasias dos Ductos Biliares/patologia , Humanos , Pessoa de Meia-Idade , RNA Mensageiro/genética , Tailândia
8.
Int J Nanomedicine ; 15: 1643-1659, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32210558

RESUMO

Purpose: Aseptic loosening is a major complication after total joint replacement. Reactive oxygen species generated by local tissue cells and liberated from implant surfaces have been suggested to cause implant failures. Surface modification of titanium (Ti)-based implants with proanthocyanidins (PAC) is a promising approach for the development of anti-oxidant defense mechanism to supplement the mechanical functions of Ti implants. In this study, a controlled PAC release system was fabricated on the surface of Ti substrates using the layer-by-layer (LBL) assembly. Materials and Methods: Polyethyleneimine (PEI) base layer was fabricated to enable layer-by-layer (LBL) deposition of hyaluronic acid/chitosan (HA/CS) multi-layers without or with the PAC. Surface topography and wettability of the fabricated HA/CS-PAC substrates were characterized by scanning electron microscopy (SEM), atomic force microscopy (AFM), Fourier-transform infrared spectroscopy (FTIR) and contact angle measurement. PAC release profiles were investigated using drug release assays. MC3T3-E1 pre-osteoblast cells were used to assess the osteo-inductive effects of HA/CS-PAC substrates under conditions H2O2-induced oxidative stress in vitro. A rat model of femoral intramedullary implantation evaluated the osseo-integration and osteo-inductive potential of the HA/CS-PAC coated Ti implants in vivo. Results: SEM, AFM, FTIR and contact angle measurements verified the successful fabrication of Ti surfaces with multi-layered HA/CS-PAC coating. Drug release assays revealed controlled and sustained release of PAC over 14 days. In vitro, cell-based assays showed high tolerability and enhanced the osteogenic potential of MC3T3-E1 cells on HA/CS-PAC substrates when under conditions of H2O2-induced oxidative stress. In vivo evaluation of femoral bone 14 days after femoral intramedullary implantation confirmed the enhanced osteo-inductive potential of the HA/CS-PAC coated Ti implants. Conclusion: Multi-layering of HA/CS-PAC coating onto Ti-based surfaces by the LBL deposition significantly enhances implant osseo-integration and promotes osteogenesis under conditions of oxidative stress. This study provides new insights for future applications in the field of joint arthroplasty.


Assuntos
Antioxidantes/farmacologia , Osteogênese/efeitos dos fármacos , Proantocianidinas/farmacologia , Próteses e Implantes , Titânio/farmacologia , Animais , Apoptose/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Quitosana/química , Liberação Controlada de Fármacos , Feminino , Ácido Hialurônico/química , Peróxido de Hidrogênio/farmacologia , Espaço Intracelular/metabolismo , Camundongos , Osseointegração/efeitos dos fármacos , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Polietilenoimina/química , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Molhabilidade , Microtomografia por Raio-X
9.
Med Sci Monit ; 26: e921087, 2020 Mar 09.
Artigo em Inglês | MEDLINE | ID: mdl-32147666

RESUMO

BACKGROUND We sought to investigate the expression of KPNA2 in bladder cancer (BC) and its relationship with prognosis, and to analyze the potential mechanism of KPNA2 in promoting BC progression. MATERIAL AND METHODS The RNA-seq data on BC from The Cancer Genome Atlas (TCGA) database were imported into R statistical software for differential analysis. The clinical data for patients with BC were screened and analyzed with R software. The survival curve was drawn with the Kaplan-Meier Plotter. The expression of KPNA2 in 4 human BC cell lines and a human bladder epithelial cell line was detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting (WB). The proliferation of BC cells was detected with Cell Counting Kit-8 (CCK8), detection of apoptosis, and flow cytometry, and the migration and invasion of BC cells were detected through Transwell assays. WB was used to detect proteins involved in the P53 pathway. RESULTS The expression of KPNA2 was higher in BC. The difference in KPNA2 expression was associated with many clinicopathological factors, and high expression of KPNA2 was associated with shorter survival time. After KPNA2 knockout, the proliferation, migration, and invasion ability decreased significantly, the cell cycle was clearly arrested in the G0/G1 phase, and the number of apoptotic cells increased. Moreover, CyclinD1, BCL2, and pro-caspase3 decreased significantly, whereas P53, P21, BAX, and cleaved-caspase3 increased significantly. The results in the overexpression group were the opposite of results in the knockdown group. CONCLUSIONS KPNA2 is an oncogenic factor that facilitates BC tumorigenicity through the P53 pathway.


Assuntos
Proteína Supressora de Tumor p53/metabolismo , Neoplasias da Bexiga Urinária/patologia , alfa Carioferinas/genética , Apoptose , Linhagem Celular Tumoral , Proliferação de Células , Técnicas de Inativação de Genes , Humanos , Estimativa de Kaplan-Meier , Prognóstico , Neoplasias da Bexiga Urinária/diagnóstico , Neoplasias da Bexiga Urinária/genética
10.
Anticancer Res ; 40(3): 1535-1542, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32132054

RESUMO

BACKGROUND/AIM: Sessile serrated polyps without dysplasia (SSPND) are characterized by crypts with serrated epithelium, albeit with irregular, corrupted shapes (CCS). PATIENTS AND METHODS: Cell proliferation was explored in the CCS from 60 SSPND and in the crypts from 12 normal colons. Sections were immuno-stained with the Ki-67 proliferation-cell (PC) marker, and with the p53 tumour-suppressor gene. RESULTS: Three predominant PC-phenotypes were found in the CCS from the 60 SSPND: 44 (73.3%) exhibited ectopic, asymmetric, randomly distributed PC-clusters, 12 (20.0%), continuous PC in one or in both slopes of the crypts, and in the remaining 4 (6.7%), single, randomly distributed PC were recorded. In contrast, the scrutiny of more than 200,000 normal colon crypts (controls) showed symmetrically aligned PC, restricted to the lower third of the crypts. p53-up-regulation in CCS was recorded in 11(18.3%) of the 60 NDSSP, but in none of the normal crypts in the 12 controls. CONCLUSION: The non-dysplastic epithelium that lines CCS in SSPND coexists with an asymmetric relocation of the PC-domains. In addition, the CCS in nearly one-fifth of the SSPND exhibited p53-up-regulated cells. Taken together, the non-dysplastic CCS epithelium in SSPND thrives with somatic mutations. The accretion of putative mutated non-dysplastic CCS might be a crucial event in the evolution of colonic SSPND towards sessile serrated adenomas.


Assuntos
Pólipos do Colo/patologia , Proliferação de Células/fisiologia , Forma Celular/fisiologia , Pólipos do Colo/genética , Pólipos do Colo/metabolismo , Genes p53 , Humanos , Imunoensaio , Fenótipo , Proteína Supressora de Tumor p53/biossíntese , Proteína Supressora de Tumor p53/genética , Regulação para Cima
11.
Gene ; 741: 144557, 2020 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-32171824

RESUMO

The MDM2 oncogene is a negative regulator of the p53 tumour suppressor. This relationship appears to have originated over a billion years ago. The human MDM2 gene encodes a variety of mRNAs with exceptionally long 3'UTRs (up to 5.7 kb); however, it was unclear whether MDM2 3'UTRs from other species are similarly long or conserved at the sequence level. Here, we report that all but one of the primate species most closely related to humans (greater and lesser apes) have similarly long 3'UTRs with high sequence similarity across their entire length. More distantly related species (Old world monkeys and new world monkeys) tend to have shorter MDM2 3'UTRs homologous to the corresponding position of the human MDM2 3'UTR while non-primate species exhibit little similarity at all. Remarkably, DNA sequences downstream of the shorter primate 3'UTRs are syntenic with distal regions in the human and other ape MDM2 3'UTRs. These homologous non-transcribed intergenic and transcribed 3'UTR-encoding regions are comprised of a variety of transposable elements, an RLP24 pseudogene and a cluster of novel repeat sequences suggestive of another unknown transposable element. Our analysis suggests that the primary difference between long and short MDM2 3'UTRs is a switch in polyA site usage to include conserved transposable elements that remain intergenic in more distantly related primates. It will be important to determine the relative contribution of these elements to post-transcriptional and translational regulation of MDM2 and hence p53-mediated tumour suppression.


Assuntos
Evolução Molecular , Genômica , Primatas/genética , Proteínas Proto-Oncogênicas c-mdm2/genética , Regiões 3' não Traduzidas/genética , Animais , Hibridização Genômica Comparativa , Elementos de DNA Transponíveis/genética , Regulação da Expressão Gênica/genética , Genoma/genética , Humanos , Poliadenilação/genética , Pseudogenes/genética , Proteína Supressora de Tumor p53/genética
12.
Nat Cell Biol ; 22(3): 321-331, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32123335

RESUMO

CRISPR-Cas9 technology has revolutionized genome editing and is applicable to the organoid field. However, precise integration of exogenous DNA sequences into human organoids is lacking robust knock-in approaches. Here, we describe CRISPR-Cas9-mediated homology-independent organoid transgenesis (CRISPR-HOT), which enables efficient generation of knock-in human organoids representing different tissues. CRISPR-HOT avoids extensive cloning and outperforms homology directed repair (HDR) in achieving precise integration of exogenous DNA sequences into desired loci, without the necessity to inactivate TP53 in untransformed cells, which was previously used to increase HDR-mediated knock-in. CRISPR-HOT was used to fluorescently tag and visualize subcellular structural molecules and to generate reporter lines for rare intestinal cell types. A double reporter-in which the mitotic spindle was labelled by endogenously tagged tubulin and the cell membrane by endogenously tagged E-cadherin-uncovered modes of human hepatocyte division. Combining tubulin tagging with TP53 knock-out revealed that TP53 is involved in controlling hepatocyte ploidy and mitotic spindle fidelity. CRISPR-HOT simplifies genome editing in human organoids.


Assuntos
Sistemas CRISPR-Cas , Edição de Genes , Técnicas de Introdução de Genes/métodos , Organoides/citologia , Hepatócitos/citologia , Hepatócitos/ultraestrutura , Humanos , Intestinos/citologia , Fígado/citologia , Organoides/ultraestrutura , Fuso Acromático/ultraestrutura , Proteína Supressora de Tumor p53/fisiologia
13.
Chem Biol Interact ; 322: 109058, 2020 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-32171848

RESUMO

Epidemiological evidence underscores alcohol consumption as a strong risk factor for multiple cancer types, with liver cancer being most commonly associated with alcohol intake. While mechanisms linking alcohol consumption to malignant tumor development are not fully understood, the likely players in ethanol-induced carcinogenesis are genotoxic stress caused by formation of acetaldehyde, increased oxidative stress, and altered nutrient metabolism, including the impairment of methyl transfer reactions. Alterations of sphingolipid metabolism and associated signaling pathways are another potential link between ethanol and cancer development. In particular, ceramides are involved in the regulation of cellular proliferation, differentiation, senescence, and apoptosis and are known to function as important regulators of malignant transformation as well as tumor progression. However, to date, the cross-talk between ceramides and alcohol in cancer disease is largely an open question and only limited data are available on this subject. Most studies linking ceramide to cancer considered liver steatosis as the underlying mechanism, which is not surprising taking into consideration that ceramide pathways are an integral part of the overall lipid metabolism. This review summarizes the latest studies pointing to ceramide as an important mediator of cancer-promoting effects of chronic alcohol consumption and underscores the necessity of understanding the role of sphingolipids and lipid signaling in response to alcohol in order to prevent and/or successfully manage diseases caused by alcohol.


Assuntos
Consumo de Bebidas Alcoólicas , Etanol/metabolismo , Neoplasias/patologia , Esfingolipídeos/metabolismo , Animais , Humanos , Neoplasias/metabolismo , Hepatopatia Gordurosa não Alcoólica/metabolismo , Hepatopatia Gordurosa não Alcoólica/patologia , Proteína Supressora de Tumor p53/metabolismo
15.
PLoS Comput Biol ; 16(2): e1007701, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-32101536

RESUMO

Tumor-specific genomic alterations allow systematic identification of genetic interactions that promote tumorigenesis and tumor vulnerabilities, offering novel strategies for development of targeted therapies for individual patients. We develop an Individualized Network-based Co-Mutation (INCM) methodology by inspecting over 2.5 million nonsynonymous somatic mutations derived from 6,789 tumor exomes across 14 cancer types from The Cancer Genome Atlas. Our INCM analysis reveals a higher genetic interaction burden on the significantly mutated genes, experimentally validated cancer genes, chromosome regulatory factors, and DNA damage repair genes, as compared to human pan-cancer essential genes identified by CRISPR-Cas9 screenings on 324 cancer cell lines. We find that genes involved in the cancer type-specific genetic subnetworks identified by INCM are significantly enriched in established cancer pathways, and the INCM-inferred putative genetic interactions are correlated with patient survival. By analyzing drug pharmacogenomics profiles from the Genomics of Drug Sensitivity in Cancer database, we show that the network-predicted putative genetic interactions (e.g., BRCA2-TP53) are significantly correlated with sensitivity/resistance of multiple therapeutic agents. We experimentally validated that afatinib has the strongest cytotoxic activity on BT474 (IC50 = 55.5 nM, BRCA2 and TP53 co-mutant) compared to MCF7 (IC50 = 7.7 µM, both BRCA2 and TP53 wild type) and MDA-MB-231 (IC50 = 7.9 µM, BRCA2 wild type but TP53 mutant). Finally, drug-target network analysis reveals several potential druggable genetic interactions by targeting tumor vulnerabilities. This study offers a powerful network-based methodology for identification of candidate therapeutic pathways that target tumor vulnerabilities and prioritization of potential pharmacogenomics biomarkers for development of personalized cancer medicine.


Assuntos
Biologia Computacional/métodos , Redes Reguladoras de Genes , Neoplasias/genética , Antineoplásicos/uso terapêutico , Proteína BRCA2/genética , Biomarcadores Tumorais , Sistemas CRISPR-Cas , Carcinogênese , Linhagem Celular Tumoral , Exoma , Testes Genéticos , Genômica , Humanos , Concentração Inibidora 50 , Modelos Teóricos , Mutação , Neoplasias/tratamento farmacológico , Farmacogenética , Medicina de Precisão , Mapeamento de Interação de Proteínas , Taxa de Sobrevida , Proteína Supressora de Tumor p53/genética
16.
Cancer Immunol Immunother ; 69(5): 835-846, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32062693

RESUMO

Tumor-associated macrophages (TAMs) are important regulators of the complex interplay between immune system and breast cancer. TAMs fuel the cancer progression and metastasis by reprogramming their specific functional phenotype in cancer settings. Therefore, it is important to clarify the mechanisms of shaping specific functional phenotype of macrophages in tumor milieu. LncRNA profiles of TAMs were identified by LncRNA microarray. Flow cytometry was used to detect the surface markers of TAMs. The co-localization among lincRNA-p21, p53 and Mouse Double Minute 2 (MDM2) was identified by FISH probe and immunofluorescence. PyVT-MMTV and BALB/c mice were used for in vivo analysis. In the present work, we found that lincRNA-p21 significantly up-regulated in 4T1 educated macrophages. LincRNA-p21 knockdown facilitated macrophage polarization into pro-inflammatory M1 in tumor microenvironment, which might be caused by MDM2 eliciting proteasome-dependent degradation to p53 and activated NF-κB and STAT3 pathway. TAMs with lincRNA-p21 knockdown induced cancer cell apoptosis, inhibited tumor cell migration and invasion. In vivo, lincRNA-p21 knockdown macrophage adoptive transfer could alleviate breast cancer progression. Our results indicated that lincRNA-p21 was a key regulator of TAMs function in tumor milieu. Our data also shed a light on novel therapeutic targets of tumors characterized by monocytes/macrophages infiltration.


Assuntos
Neoplasias da Mama/genética , Regulação Neoplásica da Expressão Gênica/imunologia , Macrófagos/imunologia , Proteínas Proto-Oncogênicas c-mdm2/genética , RNA Longo não Codificante/metabolismo , Animais , Neoplasias da Mama/imunologia , Neoplasias da Mama/patologia , Linhagem Celular Tumoral/transplante , Modelos Animais de Doenças , Feminino , Perfilação da Expressão Gênica , Técnicas de Silenciamento de Genes , Humanos , Camundongos , Análise de Sequência com Séries de Oligonucleotídeos , Cultura Primária de Células , Proteólise , Proteínas Proto-Oncogênicas c-mdm2/metabolismo , RNA Longo não Codificante/genética , Transdução de Sinais/genética , Transdução de Sinais/imunologia , Microambiente Tumoral/genética , Microambiente Tumoral/imunologia , Proteína Supressora de Tumor p53/metabolismo , Regulação para Cima
17.
Chem Biol Interact ; 317: 108972, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-32017914

RESUMO

BACKGROUND: Heart failure (HF) is an epidemic disease with increased incidence annually. It has been reported that taurine can improve cardiac function. This study investigated the cardioprotective effects of taurine in pressure-loaded HF mice and elucidated the possible mechanism. METHODS: HF models were established by transverse aortic constriction (TAC). Animals were treated with either taurine for 9 weeks and/or the SIRT1 inhibitor EX527 (5 mg/kg/day, every 2days) after TAC operation. Cardiac function and geometry were revealed by echocardiography. Myocardial hypertrophy and fibrosis were assessed using Fluorescent wheat germ agglutinin (WGA) staining and Masson's trichrome staining. Western blot and RT-PCR were performed to elucidate the expression of target proteins and genes respectively. Apoptosis in cardiomyocytes was detected by TUNEL staining. Myocardial oxidative stress was assessed by detecting the concentration of myocardial super oxidative dismutase (SOD) and malonyldialdehyde (MDA) and reactive oxygen species (ROS). Taurine concentrations and NAD+/NADH ratio were determined by taurine and NAD+/NADH assay kit. RESULTS: Taurine notably relieved cardiac dysfunction after TAC. The mechanisms were attributed to reduced myocyte hypertrophy and fibrosis, and alleviated apoptosis and oxidative stress. Meanwhile, taurine increased NAD+/NADH ratio,promoted the expression of SIRT1 and suppressed p53 acetylation. However, EX-527(inhibitor of SIRT1) decreased NAD+/NADH ratio and increased acetyl-p53 levels, and abolished the cardioprotective effects of taurine on mice subjected to TAC and increased apoptosis and oxidative stress. CONCLUSION: The mechanism responsible for cardiac-protective effects of taurine in HF induced by pressure overload is associated with the activation of the SIRT1-p53 pathway.


Assuntos
Miócitos Cardíacos/efeitos dos fármacos , Pressão , Sirtuína 1/metabolismo , Taurina/farmacologia , Proteína Supressora de Tumor p53/metabolismo , Animais , Aorta/patologia , Apoptose/efeitos dos fármacos , Carbazóis/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Ligadura , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Estresse Oxidativo , Sirtuína 1/genética , Proteína Supressora de Tumor p53/genética
18.
Anticancer Res ; 40(2): 767-777, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32014919

RESUMO

BACKGROUND/AIM: Chicken ovalbumin upstream promoter-transcription factor II (COUP-TFII) plays an important role in cancer. We examined the effect of COUP-TFII overexpression on the proliferation and invasion of the human colorectal cancer SNU-C4 cells. MATERIALS AND METHODS: SNU-C4 cells were stably transfected with COUP-TFII expression plasmid to overexpress COUP-TFII (COUP-TFII-SNU-C4 cells). Cell proliferation, colony-forming ability and transwell invasion assays were performed. To elucidate the underlying molecular mechanism of COUP-TFII action, western blot analysis, p53 shRNA transfection, and Myr-Akt transfection were performed. RESULTS: Cell proliferation and colony-forming ability were significantly inhibited in COUP-TFII-SNU-C4 cells. Western blot analyses demonstrated that while the expression of p53 and PTEN was increased, the p-Akt levels were decreased in COUP-TFII-SNU-C4 cells. Knockdown of p53 partially restored the cell proliferation, but did not reverse the inhibition of invasion. Constitutive activation of Akt via Myr-Akt transfection reversed the inhibited cell proliferation and invasion by COUP-TFII. CONCLUSION: p53 is required for the inhibition of cell proliferation, and decreased phosphorylation of Akt may mediate the inhibition of cell proliferation and invasion by COUP-TFII.


Assuntos
Fator II de Transcrição COUP/biossíntese , Neoplasias Colorretais/metabolismo , PTEN Fosfo-Hidrolase/biossíntese , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteína Supressora de Tumor p53/biossíntese , Fator II de Transcrição COUP/metabolismo , Proliferação de Células/fisiologia , Neoplasias Colorretais/patologia , Humanos , PTEN Fosfo-Hidrolase/metabolismo , Fosforilação , Transfecção , Proteína Supressora de Tumor p53/metabolismo
20.
J Cancer Res Clin Oncol ; 146(4): 861-874, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32088783

RESUMO

PURPOSE: Non-small cell lung cancer (NSCLC) is the most common type of lung cancer and ranked top in terms of incidence and mortality in men and women. Recently, improvements in treatment approaches for NSCLC have reported, but still, there is a need to devise innovative treatment strategies, especially to manage the advanced and metastatic stage of NSCLC. Aloperine (ALO), an herbal alkaloid, has exerted anti-cancer effects in many cancers. However, the use of any chemotherapeutic agents is dose limited due to possible adverse effects and drug-resistance issues. Therefore, a combination of chemotherapy with viral-based targeted gene therapy may provide a novel treatment strategy for NSCLC. METHODS/RESULTS: In this study, the results of the MTT and flow cytometry-based assays showed that Aloperine-Adbic (adenoviral vector expressing p14ARF/p53) combined treatment on NSCLC cells synergistically produced anti-proliferative effects, induced apoptosis, and arrested cell cycle at the G1 phase. Furthermore, the expression analysis suggested that the p53/p21 pathway might contribute to achieving aforesaid cytotoxic effects. The ALO-Adbic combined treatment prolonged the percent survival of NSCLC xenograft models. CONCLUSION: In conclusion, ALO-Adbic combination can produce synergistic anti-cancer effects at low doses, and may offer a more effective and less toxic new treatment strategy for NSCLC.


Assuntos
Carcinoma Pulmonar de Células não Pequenas/terapia , Terapia Genética/métodos , Neoplasias Pulmonares/terapia , Piperidinas/farmacologia , Adenoviridae/genética , Animais , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Linhagem Celular Tumoral , Terapia Combinada , Vetores Genéticos/genética , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Distribuição Aleatória , Proteína Supressora de Tumor p14ARF/genética , Proteína Supressora de Tumor p53/genética , Ensaios Antitumorais Modelo de Xenoenxerto
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