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1.
Int J Mol Sci ; 22(7)2021 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-33808426

RESUMO

Dietary methyl-donors play important roles in physiological processes catalyzed by B vitamins as coenzymes, and are used for complementary support in oncotherapy. Our hypothesis was that methyl-donors can not only assist in tolerating cancer treatment but may also directly interfere with tumor growth and proliferation. Therefore, we investigated the proposed cancer inhibitory effects of methyl-donors (in a mixture of L-methionine, choline chloride, folic acid, and vitamin B12) on MCF7 and T47D breast cancer as well as A549 and H1650 lung cancer cell lines. Indeed, methyl-donor treatment significantly reduced the proliferation in all cell lines, possibly through the downregulation of MAPK/ERK and AKT signaling. These were accompanied by the upregulation of the pro-apoptotic Bak and Bax, both in MCF7 and H1650 cells, at reduced anti-apoptotic Mcl-1 and Bcl-2 levels in MCF7 and H1650 cells, respectively. The treatment-induced downregulation of p-p53(Thr55) was likely to contribute to protecting the nuclear localization and apoptosis inducing functions of p53. The presented features are known to improve the sensitivity of cancer therapy. Therefore, these data support the hypothesis, i.e., that methyl-donors may promote apoptotic signaling by protecting p53 functions through downregulating both the MAPK/ERK and the AKT pathways both in breast and lung adenocarcinoma cell lines. Our results can emphasize the importance and benefits of the appropriate dietary supports in cancer treatments. However, further studies are required to confirm these effects without any adverse outcome in clinical settings.


Assuntos
Apoptose/fisiologia , Sistema de Sinalização das MAP Quinases/fisiologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Apoptose/efeitos dos fármacos , Mama/patologia , Neoplasias da Mama/metabolismo , Linhagem Celular Tumoral/metabolismo , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Colina/farmacologia , Ácido Fólico/farmacologia , Humanos , Pulmão/patologia , Neoplasias Pulmonares/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Metionina/farmacologia , Metilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-akt/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Vitamina B 12/farmacologia
2.
Mol Med Rep ; 23(5)2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33760119

RESUMO

Aberrant microRNA (miRNA/miR) expression plays an important role in the pathogenesis of nasopharyngeal carcinoma (NPC). In the present study, the role and underlying molecular mechanism of miR­301a­3p in NPC cells were determined. It was observed that miR­301a­3p upregulation promoted NPC cell proliferation, migration, invasion and epithelial­mesenchymal transition in vitro, whereas its downregulation resulted in the opposite effect. B­cell translocation gene 1 (BTG1) mRNA was identified as the novel target of miR­301a­3p. BTG1 overexpression partially attenuated miR­301a­3p­induced increase in cell proliferation and invasion. miR­301a­3p can be transferred by exosomes and positively regulate the proliferation and invasion of NPC cells. Altogether, the present study highlights that exosomal miR­301a­3p can promote NPC cell proliferation and invasion by repressing BTG1, thereby resulting in the development of NPC.


Assuntos
Proliferação de Células/genética , MicroRNAs/genética , Carcinoma Nasofaríngeo/genética , Proteínas de Neoplasias/genética , Linhagem Celular Tumoral/metabolismo , Movimento Celular/genética , Transição Epitelial-Mesenquimal/genética , Exossomos/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Carcinoma Nasofaríngeo/patologia , Invasividade Neoplásica/genética , Invasividade Neoplásica/patologia , RNA Mensageiro/genética
3.
Medicine (Baltimore) ; 99(27): e20944, 2020 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-32629699

RESUMO

BACKGROUND: Colorectal cancer is the second commonly seen cancer around the world and accounts for 13% of all human cancers. Among them, 25% of all case were diagnosed with metastasis and 50% occurs metastasis during the development of disease. Cetuximab is a chimeric monoclonal antibody against epidermal growth factor receptor, and is used for treatment of metastatic colorectal cancer alone or combined with chemotherapy or radiation therapy. Integrin-beta 1 (ITGB1), which is also known as CD29, and plays an important role in development of malignant cancers. However, the effect of ITGB1 in promoting the anti-tumor effect of cetuximab is not fully understand. METHODS: The model of ITGB1 inhibition and overexpression was firstly constructed in LS174T cells, and the viability of cells in each group was detected using CCK-8 assay. The expression of key factors in tumor formation process at transcription level was detected using real-time quantitative polymerase chain reaction method. The expression of key proteins in metastasis process, cell apoptosis and activation of Ras/Raf/MEK signaling pathway was detected using western blotting analysis. And the concentration of key factors of in tumor formation process in cultured medium of LS174T cells were detected using enzyme-linked immunosorbent assay method. RESULTS: We found that cetuximab could inhibit the proliferation of LS174T cells, and inhibition of ITGB1 enhanced this effect while overexpression of ITGB1 reduced this effect. We further found that cetuximab could inhibit the expression and secretion of extracellular matrix degradation related molecules in cultured medium and transcription level. Besides, we also found that the expression of key factors in angiogenesis and extracellular matrix degradation related proteins were also reduced after cetuximab treatment. These effects might be mediated by Ras/Raf/MAPK signaling pathway and enhanced after inhibition of ITGB1 expression. CONCLUSION: Inhibition of ITGB1 might be a new therapeutic method in colorectal cancer.


Assuntos
Antineoplásicos Imunológicos/farmacologia , Cetuximab/farmacologia , Neoplasias Colorretais/tratamento farmacológico , Integrina beta1/efeitos dos fármacos , Antineoplásicos Imunológicos/uso terapêutico , Linhagem Celular Tumoral/efeitos dos fármacos , Linhagem Celular Tumoral/metabolismo , Cetuximab/uso terapêutico , Neoplasias Colorretais/patologia , Humanos , Integrina beta1/metabolismo
4.
Arch Biochem Biophys ; 689: 108462, 2020 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-32590068

RESUMO

Silver nanoparticles (AgNP) emerged as a promising reagent for cancer therapy with oxidative stress implicated in the toxicity. Meanwhile, studies reported cold atmospheric plasma (CAP) generation of reactive oxygen and nitrogen species has selectivity towards cancer cells. Gold nanoparticles display synergistic cytotoxicity when combined with CAP against cancer cells but there is a paucity of information using AgNP, prompting to investigate the combined effects of CAP using dielectric barrier discharge system (voltage of 75 kV, current is 62.5 mA, duty cycle of 7.5kVA and input frequency of 50-60Hz) and 10 nm PVA-coated AgNP using U373MG Glioblastoma Multiforme cells. Cytotoxicity in U373MG cells was >100-fold greater when treated with both CAP and PVA-AgNP compared with either therapy alone (IC50 of 4.30 µg/mL with PVA-AgNP alone compared with 0.07 µg/mL after 25s CAP and 0.01 µg/mL 40s CAP). Combined cytotoxicity was ROS-dependent and was prevented using N-Acetyl Cysteine. A novel darkfield spectral imaging method investigated and quantified AgNP uptake in cells determining significantly enhanced uptake, aggregation and subcellular accumulation following CAP treatment, which was confirmed and quantified using atomic absorption spectroscopy. The results indicate that CAP decreases nanoparticle size, decreases surface charge distribution of AgNP and induces uptake, aggregation and enhanced cytotoxicity in vitro.


Assuntos
Neoplasias Encefálicas/terapia , Glioblastoma/terapia , Gases em Plasma/farmacologia , Prata/farmacologia , Neoplasias Encefálicas/metabolismo , Linhagem Celular Tumoral/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Glioblastoma/metabolismo , Humanos , Nanopartículas Metálicas/análise , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Prata/farmacocinética
5.
Biol Res ; 53(1): 14, 2020 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-32293550

RESUMO

BACKGROUND: Previous studies have shown that long noncoding RNA (lncRNA) LINC00483 was aberrantly expressed in human cancers, including gastric cancer. However, the regulatory mechanism of this lncRNA in gastric cancer remains largely unknown. The present study aimed to investigate the effect of LINC00483 on gastric cancer development and explore the potential regulatory network of LINC00483/microRNA (miR)-490-3p/mitogen-activated protein kinase 1 (MAPK1). METHODS: Thirty patients with gastric cancer were recruited for tissues collection. The expression levels of LINC00483, miR-490-3p and MAPK1 were detected by quantitative real-time polymerase chain reaction or western blot. Cell viability, apoptosis, migration and invasion were determined by MTT, flow cytometry, transwell assays and western blot, respectively. The target association between miR-490-3p and LINC00483 or MAPK1 was confirmed by luciferase reporter assay. Xenograft model was established to assess the function of LINC00483 in vivo. RESULTS: LINC00483 and MAPK1 levels were increased in gastric cancer tissues and cells. Knockdown of LINC00483 or MAPK1 inhibited cells viability, migration and invasion but promoted apoptosis in gastric cancer cells. Moreover, MAPK1 overexpression attenuated the effect of LINC00483 knockdown on gastric cancer development. LINC00483 could increase MAPK1 expression by competitively sponging miR-490-3p. miR-490-3p overexpression suppressed gastric cancer development, which was abated by introduction of LINC00483. Besides, inhibition of LINC00483 decreased xenograft tumor growth by regulating miR-490-3p/MAPK1 axis. CONCLUSION: Knockdown of LINC00483 inhibited gastric cancer development in vitro and in vivo by increasing miR-490-3p and decreasing MAPK1, elucidating a novel mechanism for understanding the development of gastric cancer.


Assuntos
MicroRNAs/metabolismo , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , RNA Longo não Codificante/metabolismo , Neoplasias Gástricas/metabolismo , Animais , Apoptose , Carcinogênese/metabolismo , Linhagem Celular Tumoral/metabolismo , Movimento Celular , Sobrevivência Celular , Células Epiteliais/metabolismo , Regulação Neoplásica da Expressão Gênica , Humanos , Luciferases/metabolismo , Masculino , Camundongos Endogâmicos BALB C , MicroRNAs/genética , RNA Longo não Codificante/genética , Neoplasias Gástricas/genética , Ensaios Antitumorais Modelo de Xenoenxerto
6.
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
7.
Biomed Res Int ; 2020: 5393041, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32149115

RESUMO

Resveratrol (Resv) offers health benefits in cancer and has been reported to modulate important enzymes of lipid metabolism. Studies of its effects on lipid composition in different subtypes of breast-cancer cells are scarce. Thus, we investigated the alterations in phospholipids (PL), fatty acids (FA), and lipid metabolism enzymes in two breast-cancer cell lines after Resv treatment. MCF-7 and MDA-MB-231 cells were treated with 80 and 200 µM of Resv, respectively, for 24 hours. We analyzed PL with radiolabeled inorganic phosphate (32Pi) by thin-layer chromatography, FA by gas chromatography-mass spectrometry, and lipid metabolism enzymes (DGAT2, FAS, ρACCß, pAMPKα, and AMPK) by Western blot. Resv treated MDA-MB-231 phospholipids showed a reduction in phosphatidylcholine (63%) and phosphatidylethanolamine (35%). We observed an increase in eicosapentaenoic acid (EPA) (73%) and docosahexaenoic acid (DHA) (65%) in MCF-7 cells after Resv treatment. Interestingly, the same treatment caused 50% and 90% increases in EPA and DHA, respectively, in MDA-MB-231 cells. In MCF-7 cells, Resv increased the expression of ρACCß (3.3-fold) and AMPKα/ρAMPKα (1.5-fold) and in MDA-MB-231 cells it inhibited the expression of ρACCß (111.8-fold) and AMPKα/ρAMPKα (1.2 fold). Our results show that Resv modified PL and saturated and unsaturated FA especially in MDA-MB-231 cells, and open new perspectives to the understanding of the reported anticancer effect of Resv on these cells.


Assuntos
Neoplasias da Mama/tratamento farmacológico , Metabolismo dos Lipídeos/efeitos dos fármacos , Resveratrol/farmacologia , Proteínas Quinases Ativadas por AMP/metabolismo , Neoplasias da Mama/metabolismo , Linhagem Celular Tumoral/efeitos dos fármacos , Linhagem Celular Tumoral/metabolismo , Ácidos Docosa-Hexaenoicos/metabolismo , Ácido Eicosapentaenoico/análogos & derivados , Ácidos Graxos/metabolismo , Ácidos Graxos Insaturados , Feminino , Humanos , Lipídeos/análise , Células MCF-7 , Fosfatidilcolinas/metabolismo , Fosfatidiletanolaminas/metabolismo , Fosfolipídeos/metabolismo , Resveratrol/uso terapêutico
8.
J Cardiothorac Surg ; 15(1): 18, 2020 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-31931858

RESUMO

INTRODUCTION: Lung cancer is the leading causes of cancer-related deaths globally. The most frequent histologic type of lung cancer is non-small-cell lung cancer (NSCLC). NSCLC often undergo epithelial-mesenchymal transition (EMT). The components that control this process are thus promising therapeutic targets. MATERIALS AND METHODS: Gli/EMT protein expression levels were examined by western blot in paired NSCLC patient tissues and NSCLC cell lines. Functional analyses were performed to investigate SHH/Gli signaling and EMT in NSCLC cell lines. MTS cell viability, luciferase reporter, and western blot assays were performed to analyze pathway activity, while wound healing and transwell assays were executed to measure cell migration and invasion. RESULTS: Higher Gli1 expressions were detected in tumor samples than in paired normal tissues. Differential expression of EMT biomarkers and activation of p-AKT were observed in tumor tissues. N-Shh stimulation of cells significantly increased reporter activity in NSCLC cell lines, while Gli-i treatment of transfected cells showed less relative reporter activity. When subjected to both Gli-i and N-Shh treatment, NSCLC cell lines continued to demonstrate decreased Gli transcriptional activity. Gli inhibition is associated with decreased expression level of p-AKT, N-cadherin and Vimentin. Knockdown of both Gli1 and Gli2 showed decreased EMT, migrative and invasive ability. Cells stimulated by N-Shh demonstrated greater mobility. In addition, AKT-i treated cells also demonstrated inhibited EMT activity. CONCLUSIONS: This study provides evidence for aberrant upregulation of the Gli signaling pathway and a strong association between expression of Gli versus AKT and EMT markers in NSCLC.


Assuntos
Carcinoma Pulmonar de Células não Pequenas/metabolismo , Transição Epitelial-Mesenquimal/fisiologia , Neoplasias Pulmonares/metabolismo , Proteína GLI1 em Dedos de Zinco/fisiologia , Biomarcadores Tumorais/metabolismo , Caderinas/metabolismo , Linhagem Celular Tumoral/metabolismo , Movimento Celular/efeitos dos fármacos , Proteínas Hedgehog/fisiologia , Humanos , Transdução de Sinais/fisiologia , Proteína GLI1 em Dedos de Zinco/metabolismo
9.
Biol. Res ; 53: 13, 2020. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1100919

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
Humanos , Animais , Masculino , Pessoa de Meia-Idade , Antígenos Glicosídicos Associados a Tumores/genética , Índios Sul-Americanos/genética , Neoplasias da Vesícula Biliar/genética , Líquido Ascítico/metabolismo , Células Tumorais Cultivadas , Testes de Carcinogenicidade , Chile , Impressões Digitais de DNA , Proteína Supressora de Tumor p53/genética , Cisplatino/farmacologia , Camundongos Endogâmicos NOD , Células Clonais/efeitos dos fármacos , Células Clonais/metabolismo , Análise de Sequência de RNA , Receptor ErbB-2/genética , Genes erbB-2/genética , Perfilação da Expressão Gênica , Linhagem Celular Tumoral/efeitos dos fármacos , Linhagem Celular Tumoral/metabolismo , Desoxicitidina/análogos & derivados , Desoxicitidina/farmacologia , Células Epiteliais/metabolismo , Queratina-19/genética , Queratina-7/genética , Carcinogênese/genética , Neoplasias da Vesícula Biliar/metabolismo , Antineoplásicos/farmacologia
10.
Biol. Res ; 53: 14, 2020. graf
Artigo em Inglês | LILACS | ID: biblio-1100920

RESUMO

BACKGROUND: Previous studies have shown that long noncoding RNA (IncRNA) LINC00483 was aberrantly expressed in human cancers, including gastric cancer. However, the regulatory mechanism of this IncRNA in gastric cancer remains largely unknown. The present study aimed to investigate the effect of LINC00483 on gastric cancer development and explore the potential regulatory network of LINC00483/microRNA (miR)-490-3p/mitogen-activated protein kinase 1 (MAPK1). METHODS: Thirty patients with gastric cancer were recruited for tissues collection. The expression levels of LINC00483, miR-490-3p and MAPK1 were detected by quantitative real-time polymerase chain reaction or western blot. Cell viability, apoptosis, migration and invasion were determined by MTT, flow cytometry, transwell assays and western blot, respectively. The target association between miR-490-3p and LINC00483 or MAPK1 was confirmed by luciferase reporter assay. Xenograft model was established to assess the function of LINC00483 in vivo. RESULTS: LINC00483 and MAPK1 levels were increased in gastric cancer tissues and cells. Knockdown of LINC00483 or MAPK1 inhibited cells viability, migration and invasion but promoted apoptosis in gastric cancer cells. Moreover, MAPK1 overexpression attenuated the effect of LINC00483 knockdown on gastric cancer development. LINC00483 could increase MAPK1 expression by competitively sponging miR-490-3p. miR-490-3p overexpression suppressed gastric cancer development, which was abated by introduction of LINC00483. Besides, inhibition of LINC00483 decreased xenograft tumor growth by regulating miR-490-3p/MAPK1 axis. CONCLUSION: Knockdown of LINC00483 inhibited gastric cancer development in vitro and in vivo by increasing miR- 490-3p and decreasing MAPK1, elucidating a novel mechanism for understanding the development of gastric cancer.


Assuntos
Humanos , Animais , Masculino , Neoplasias Gástricas/metabolismo , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , MicroRNAs/metabolismo , RNA Longo não Codificante/metabolismo , Neoplasias Gástricas/genética , Regulação Neoplásica da Expressão Gênica , Movimento Celular , Sobrevivência Celular , Apoptose , Ensaios Antitumorais Modelo de Xenoenxerto , MicroRNAs/genética , Linhagem Celular Tumoral/metabolismo , Células Epiteliais/metabolismo , RNA Longo não Codificante/genética , Carcinogênese/metabolismo , Luciferases/metabolismo , Camundongos Endogâmicos BALB C
11.
Life Sci Alliance ; 2(6)2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31792061

RESUMO

Colorectal cancer (CRC) is a common cancer with a high mortality rate and a rising incidence rate in the developed world. Molecular profiling techniques have been used to better understand the variability between tumors and disease models such as cell lines. To maximize the translatability and clinical relevance of in vitro studies, the selection of optimal cancer models is imperative. We have developed a deep learning-based method to measure the similarity between CRC tumors and disease models such as cancer cell lines. Our method efficiently leverages multiomics data sets containing copy number alterations, gene expression, and point mutations and learns latent factors that describe data in lower dimensions. These latent factors represent the patterns that are clinically relevant and explain the variability of molecular profiles across tumors and cell lines. Using these, we propose refined CRC subtypes and provide best-matching cell lines to different subtypes. These findings are relevant to patient stratification and selection of cell lines for early-stage drug discovery pipelines, biomarker discovery, and target identification.


Assuntos
Linhagem Celular Tumoral/classificação , Neoplasias Colorretais/classificação , Aprendizado Profundo , Biomarcadores Tumorais/genética , Linhagem Celular Tumoral/metabolismo , Linhagem Celular Tumoral/patologia , Neoplasias Colorretais/genética , Neoplasias Colorretais/patologia , Variações do Número de Cópias de DNA/genética , Perfilação da Expressão Gênica/métodos , Humanos , Aprendizado de Máquina , Modelos Biológicos , Mutação/genética , Proteínas de Neoplasias/genética , Análise de Sequência de DNA/métodos
12.
Med Sci Monit ; 25: 9829-9835, 2019 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-31864232

RESUMO

BACKGROUND This study aimed to investigate the expression profile of the phosphatase and tensin homolog (PTEN) gene, the cadherin genes, CDH1 and CDH2, and the cell membrane glycoprotein, CD133, in the Ishikawa human endometrial adenocarcinoma cell line. MATERIAL AND METHODS The Ishikawa endometrial carcinoma cell groups included cells transfected with the pLVX-puro lentiviral expression vector (the Ishikawa-puro group) and cells transfected with the pLVX-puro-PTEN lentiviral expression vector (the Ishikawa-PTEN group). The mRNA expression of the cadherin genes, CDH1 and CDH2, was detected by quantitative reverse transcription-polymerase chain reaction (RT-qPCR). The expression levels of the transmembrane glycoprotein CD133, a cancer stem cell marker, was detected by flow cytometry. RESULTS The expression of CDH1 and CDH2 mRNA in the Ishikawa-PTEN cells was lower than in the control cells. CD133 expression was lower in the Ishikawa-PTEN cells compared with the control cells. CONCLUSIONS This in vitro study showed that in Ishikawa endometrial carcinoma cells, downregulation of PTEN was associated with the expression of the CDH1 and CDH2 genes and upregulated expression of the cell membrane glycoprotein, CD133, which are associated with epithelial-mesenchymal transition (EMT) in malignancy. These findings support the need for further studies to investigate the potential role of PTEN in invasion and metastasis in endometrial carcinoma.


Assuntos
Antígeno AC133/biossíntese , Antígenos CD/biossíntese , Caderinas/biossíntese , Neoplasias do Endométrio/genética , Neoplasias do Endométrio/metabolismo , PTEN Fosfo-Hidrolase/biossíntese , Antígeno AC133/genética , Adenocarcinoma/genética , Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Antígenos CD/genética , Apoptose/fisiologia , Caderinas/genética , Linhagem Celular Tumoral/metabolismo , Neoplasias do Endométrio/patologia , Transição Epitelial-Mesenquimal , Feminino , Expressão Gênica , Humanos , PTEN Fosfo-Hidrolase/genética , Tensinas/metabolismo , Transcriptoma
13.
Sci Rep ; 9(1): 16647, 2019 11 12.
Artigo em Inglês | MEDLINE | ID: mdl-31719636

RESUMO

The present state of cancer chemotherapy is unsatisfactory. New anticancer drugs that marginally improve the survival of patients continue to be developed at an unsustainably high cost. The study aimed to elucidate the effects of insulin (INS), an inexpensive drug with a convincing safety profile, on the susceptibility of colon cancer to chemotherapeutic agents: 5-fluorouracil (FU), oxaliplatin (OXA), irinotecan (IRI), cyclophosphamide (CPA) and docetaxel (DOC). To examine the effects of insulin on cell viability and apoptosis, we performed an in vitro analysis on colon cancer cell lines Caco-2 and SW480. To verify the results, we performed in vivo analysis on mice bearing MC38 colon tumors. To assess the underlying mechanism of the therapy, we examined the mRNA expression of pathways related to the signaling downstream of insulin receptors (INSR). Moreover, we performed Western blotting to confirm expression patterns derived from the genetic analysis. For the quantification of circulating tumor cells in the peripheral blood, we used the maintrac method. The results of our study show that insulin-pretreated colon cancer cells are significantly more susceptible to commonly used chemotherapeutics. The apoptosis ratio was also enhanced when INS was administered complementary to the examined drugs. The in vivo study showed that the combination of INS and FU resulted in significant inhibition of tumor growth and reduction of the number of circulating tumor cells. This combination caused a significant downregulation of the key signaling substrates downstream of INSR. The results indicate that the downregulation of PIK3CA (phosphatidylinositol 3-kinase catalytic subunit alpha), which plays a critical role in cell signaling and GRB2 (growth factor receptor-bound protein 2), a regulator of cell proliferation and differentiation may be responsible for the sensitizing effect of INS. These findings were confirmed at protein levels by Western blotting. In conclusion, these results suggest that INS might be potentially applied to clinical use to enhance the therapeutic effectiveness of chemotherapeutic drugs. The findings may become a platform for the future development of new and inexpensive strategies for the clinical chemotherapy of tumors.


Assuntos
Antineoplásicos/uso terapêutico , Classe I de Fosfatidilinositol 3-Quinases/antagonistas & inibidores , Neoplasias Colorretais/tratamento farmacológico , Proteína Adaptadora GRB2/antagonistas & inibidores , Insulina/farmacologia , Animais , Western Blotting , Células CACO-2/efeitos dos fármacos , Células CACO-2/metabolismo , Linhagem Celular Tumoral/efeitos dos fármacos , Linhagem Celular Tumoral/metabolismo , Classe I de Fosfatidilinositol 3-Quinases/metabolismo , Neoplasias Colorretais/metabolismo , Ciclofosfamida/uso terapêutico , Docetaxel/uso terapêutico , Regulação para Baixo/efeitos dos fármacos , Sinergismo Farmacológico , Feminino , Fluoruracila/uso terapêutico , Proteína Adaptadora GRB2/metabolismo , Humanos , Irinotecano/uso terapêutico , Camundongos , Camundongos Endogâmicos C57BL , Transplante de Neoplasias , Células Neoplásicas Circulantes/efeitos dos fármacos , Células Neoplásicas Circulantes/metabolismo , Oxaliplatina/uso terapêutico
14.
Eur Rev Med Pharmacol Sci ; 23(18): 7863-7873, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31599411

RESUMO

OBJECTIVE: Hepatocellular carcinoma (HCC) is a hypervascularized tumor. Aberrant angiogenesis is the main cause, which results in cancer growth and progression. It has been showed that microRNA-302 cluster (miR-302) may be associated with angiogenesis. Here, we aimed to identify the role of miR-302a/b/c in the regulation of cell angiogenesis in HCC. PATIENTS AND METHODS: MRNA expression of miR-302a/b/c and MACC1 was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The protein of MACC1 was measured using Western blot. Cells proliferation, migration, and invasion abilities were investigated via Cell Counting Kit-8 (CCK-8) assay or transwell assay, respectively. Tube formatting assays were used to explore the tube formation capacity. The interaction among miR-302a/b/c was analyzed by luciferase assay. RESULTS: The expression of miR-302a/b/c was greatly reduced while MACC1 expression, whether mRNA or protein was conspicuously elevated in HCC tissues and cells. Then, functional experiment results showed miR-302a/b/c overexpression and MACC1 down-regulation inhibited the proliferation, migration, invasion ability, and tube formation capacity of HUVECs. In addition, we detected that miR-302a/b/c directly targeted MACC1 and suppressed MACC1 expression, and miR-302a/b/c could suppress tumor angiogenesis in HCC by targeting MACC1. CONCLUSIONS: MiR-302a/b/c may function as a potential suppressor of tumor angiogenesis in HCC by targeting MACC1, indicating a promising target for HCC therapy.


Assuntos
Carcinoma Hepatocelular/genética , Neoplasias Hepáticas/patologia , MicroRNAs/genética , Neovascularização Patológica/genética , Carcinoma Hepatocelular/mortalidade , Estudos de Casos e Controles , Linhagem Celular Tumoral/metabolismo , Movimento Celular/genética , Proliferação de Células/genética , China/epidemiologia , Regulação para Baixo , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Invasividade Neoplásica/genética , Neovascularização Patológica/patologia , Sincalida/metabolismo , Transativadores/metabolismo
15.
Eur Rev Med Pharmacol Sci ; 23(18): 7874-7883, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31599412

RESUMO

OBJECTIVE: The aim of this study was to figure out the effect of microRNA-217 on the proliferation of hepatocellular carcinoma (HCC) cells, and to explore its influence on KLF5 expression and the underlying regulatory mechanisms. PATIENTS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was used to detect the expression of microRNA-217 in tumor tissues and paracancerous tissues of 60 patients with HCC. Meanwhile, the relationship between microRNA-217 expression and HCC pathological parameters was analyzed. In HCC cell lines, including HepG2 and Bel-7402, negative control group (NC) and microRNA-217 overexpression group were set up, and qRT-PCR was performed to further verify their transfection efficiency. In addition, Cell Counting Kit-8 (CCK-8), 5-Ethynyl-2'-deoxyuridine (EdU) assay were performed to analyze the effect of microRNA-217 on the biological function of HCC cells. Finally, the potential mechanism of KLF5, the downstream gene of microRNA-217, was explored using bioinformatics analysis and cell recovery experiments. RESULTS: QRT-PCR results showed that microRNA-217 level in tumor tissues of HCC patients was conspicuously lower than that in adjacent tissues, and the difference was statistically significant. Compared with patients with high expression of microRNA-217, the pathological stage was higher and the overall survival rate was lower in patients with low expression. Compared with the NC group, the cell proliferation ability of the microRNA-217 mimics group was conspicuously decreased. Subsequently, in the HCC cell line and tissue verification, the expression of KLF5 was found remarkably increased, and microRNA-217 exhibited a negative correlation with KLF5 level. In addition, the overexpression of microRNA-217 conspicuously reduced the protein expression of CD31, Ki-67, c-Myc, MMP-2, and MMP-9. In cell recovery experiment, it was found that the overexpression of KLF5 could counteract the effect of microRNA-217 mimics on the cell proliferation of HCC, thereby inhibiting the malignant progression of this disease. CONCLUSIONS: The above studies demonstrated that microRNA-217 was markedly associated with the pathological stage and poor prognosis of HCC, and could inhibit the malignant progression of this disease. In addition, our investigation has showed that microRNA-217 might be capable of inhibiting cell proliferation of HCC via regulating KLF5.


Assuntos
Carcinoma Hepatocelular/genética , Fatores de Transcrição Kruppel-Like/metabolismo , Neoplasias Hepáticas/patologia , MicroRNAs/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma Hepatocelular/mortalidade , Estudos de Casos e Controles , Linhagem Celular Tumoral/metabolismo , Proliferação de Células/genética , China/epidemiologia , Biologia Computacional/métodos , Progressão da Doença , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias/métodos , Prognóstico , Sincalida/metabolismo , Transfecção
16.
Eur Rev Med Pharmacol Sci ; 23(18): 7892-7898, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31599414

RESUMO

OBJECTIVE: FoxO3a is a well-defined tumor suppressor gene in the forkhead transcription factor O subfamily (FoxO), and its reduction is related to the occurrence of various tumors. It was found that miR-223 expression is abnormally elevated in pancreatic cancer tissues. Bioinformatics analysis revealed a targeted complementary binding relationship between miR-223 and FoxO3a. This study explored whether miR-155 regulates the expression of FoxO3a and affects the proliferation, apoptosis, and cisplatin (CDDP) resistance of oral cancer cells. MATERIALS AND METHODS: Dual-Luciferase reporter gene assay validated the targeted relationship between miR-223 and FoxO3a. The CDDP-resistant pancreatic cancer cell line BXPC3/CDDP was established, and the expressions of miR-223 and FoxO3a were compared. BXPC3/CDDP cells were divided into miR-NC group and miR-223 inhibitor group. QRT-PCR was adopted to test miR-223 and FoxO3a mRNA expressions. Western blot was performed to determine FoxO3a protein expression. Cell apoptosis was detected by flow cytometry and cell proliferation was detected by EdU staining. RESULTS: There was a targeted regulatory relationship between miR-223 and FoxO3a mRNA. The expression of miR-223 was significantly higher, while the expression of FoxO3a mRNA and protein was significantly lower in BXPC3/CDDP cells than that in BXPC3 cells. Cell Counting Kit-8 (CCK-8) experiments showed that the same concentration of CDDP exhibited significantly lower proliferation inhibition in BXPC3/CDDP cells than BXPC3 cells. Compared with miR-NC group, transfection of miR-223 inhibitor significantly increased the expression of FoxO3a in BXPC3/CDDP cells, which significantly attenuated cell proliferation and enhanced apoptosis in CDDP-treated cells. CONCLUSIONS: Increased expression of miR-233 was associated with CDDP resistance in pancreatic cancer cells. Inhibition of miR-223 expression upregulated FoxO3a expression, restrained pancreatic cancer cell proliferation, promoted cell apoptosis, and enhanced CDDP sensitivity in pancreatic cancer cells.


Assuntos
Antineoplásicos/farmacologia , Cisplatino/farmacologia , Proteína Forkhead Box O3/efeitos dos fármacos , MicroRNAs/genética , Neoplasias Pancreáticas/genética , Apoptose/efeitos dos fármacos , Proteínas Reguladoras de Apoptose/metabolismo , Estudos de Casos e Controles , Linhagem Celular Tumoral/efeitos dos fármacos , Linhagem Celular Tumoral/metabolismo , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/genética , Biologia Computacional/métodos , Resistencia a Medicamentos Antineoplásicos/genética , Proteína Forkhead Box O3/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Neoplasias Bucais/genética , Neoplasias Pancreáticas/mortalidade , Sincalida/metabolismo , Análise de Sobrevida , Regulação para Cima/efeitos dos fármacos
17.
Eur Rev Med Pharmacol Sci ; 23(18): 7884-7891, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31599450

RESUMO

OBJECTIVE: MiR-199 expression is associated with liver cancer. Bioinformatics analysis revealed that miR-199 has a complementary binding site to the 3'-UTR region of Snail mRNA. This study investigated whether miR-199 plays a role in regulating Snail expression and affecting epithelial-mesenchymal transition (EMT) and invasion of hepatoma cells. PATIENTS AND METHODS: The Dual-Luciferase reporter gene assay validated the targeted regulation between miR-199 and Snail. QRT-PCR was used to detect and compare the expression of miR-199 and Snail mRNA in human normal liver HL7702 cells, low metastatic MHCC97L cells, and high metastatic MHCC97H cells. MHCC97H cells were cultured in vitro and divided into two groups: miR-NC group and the miR-199 mimic group followed by the analysis of the expression of Snail, E-cadherin, and N-cadherin, as well as cell invasion ability by transwell assay. RESULTS: There was a targeted regulatory relationship between miR-199 and Snail mRNA. Compared with HL7702 cells, miR-199 expression was significantly decreased, and Snail expression was significantly increased in MHCC97L and MHCC97H cells, with more changes being observed in high metastatic MHCC97H cells. The transfection of miR-199 mimic significantly downregulated the expression of Snail and N-cadherin in MHCC97H cells, increased E-cadherin expression, inhibited the cell's EMT process, and invasion. CONCLUSIONS: The decrease of miR-199 expression plays a role in upregulating the expression of Snail and promoting EMT and invasion of hepatocarcinoma cells. The increase of the expression of miR-199 can inhibit the expression of Snail and inhibit the EMT process and invasion ability of hepatoma cells.


Assuntos
Transição Epitelial-Mesenquimal/efeitos dos fármacos , Neoplasias Hepáticas/patologia , MicroRNAs/farmacologia , Regiões 3' não Traduzidas/genética , Caderinas/metabolismo , Carcinoma Hepatocelular/genética , Estudos de Casos e Controles , Linhagem Celular Tumoral/metabolismo , Linhagem Celular Tumoral/patologia , China/epidemiologia , Biologia Computacional/métodos , Regulação para Baixo , Transição Epitelial-Mesenquimal/genética , Regulação Neoplásica da Expressão Gênica/genética , Humanos , MicroRNAs/genética , Invasividade Neoplásica/genética , Fatores de Transcrição da Família Snail/genética , Regulação para Cima
18.
BMC Vet Res ; 15(1): 357, 2019 Oct 22.
Artigo em Inglês | MEDLINE | ID: mdl-31640712

RESUMO

BACKGROUND: Canine and human osteosarcomas (OS) are notably similar and have a high rate of metastasis. There is a poor understanding of the tumor development process, predisposing causes, and varying levels of aggression among different cell lines. By characterizing newly developed canine osteosarcoma cell lines, treatments for people and pets can be developed. Of the seven subtypes of OS, three are represented in this group: osteoblastic (the most common), fibroblastic, and giant cell variant. To our knowledge, there are no other giant cell variant canine OS cell lines in the published literature and only one canine fibroblastic osteosarcoma cell line. Understanding the differences between the histologic subtypes in dogs will help to guide comparative research. RESULTS: Alkaline phosphatase expression was ubiquitous in all cell lines tested and invasiveness was variable between the cell lines tested. Invasiveness and oxidative damage were not correlated with in vivo growth rates, where TOT grew the fastest and had the higher percentage of mice with metastatic lesions. TOL was determined to be the most chemo-resistant during cisplatin chemotherapy while TOM was the most chemo-sensitive. CONCLUSIONS: Further comparisons and studies using these cell lines may identify a variety of characteristics valuable for understanding the disease process and developing treatments for osteosarcoma in both species. Some of this data was presented as a poster by KMF at the August 5th, 2017 National Veterinary Scholars Program in Bethesda, MA. Characterization of 5 newly generated canine osteosarcoma cell lines. Kelli Franks, Tasha Miller, Heather Wilson-Robles.


Assuntos
Linhagem Celular Tumoral , Doenças do Cão/metabolismo , Osteossarcoma/veterinária , Adipogenia , Fosfatase Alcalina/biossíntese , Animais , Antineoplásicos/farmacologia , Linhagem Celular Tumoral/citologia , Linhagem Celular Tumoral/efeitos dos fármacos , Linhagem Celular Tumoral/metabolismo , Proliferação de Células , Condrogênese , Cisplatino/farmacologia , Meios de Cultura , Doenças do Cão/patologia , Cães , Feminino , Xenoenxertos/metabolismo , Camundongos , Invasividade Neoplásica , Transplante de Neoplasias , Osteogênese , Osteossarcoma/metabolismo
19.
Oncol Rep ; 42(6): 2826-2835, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31638243

RESUMO

In the majority of human tumors, downregulation of major histocompatibility complex class I (MHC­I) expression contributes to the escape from the host immune system and resistance to immunotherapy. Relevant animal models are therefore needed to enhance the efficacy of cancer immunotherapy. As loss of ß­2 microglobulin expression results in irreversible downregulation of surface MHC­I molecules in various human tumors, the ß­2 microglobulin gene (B2m) was deactivated in a mouse oncogenic TC­1 cell line and a TC­1/dB2m cell line that was negative for surface MHC­I expression was derived. Following stimulation with interferon γ, MHC­I heavy chains, particularly the H­2Db molecules, were found to be expressed at low levels on the cell surface, but without ß­2 microglobulin. B2m deactivation in TC­1/dB2m cells led to reduced proliferation and tumor growth. These cells were insensitive to DNA vaccination and only weakly responsive to combined immunotherapy with a DNA vaccine and the ODN1826 adjuvant. In vivo depletion demonstrated that NK1.1+ cells were involved in both reduced tumor growth and an antitumor effect of immunotherapy. The number of immune cells infiltrating TC­1/dB2m­induced tumors was comparable with that in tumors developing from TC­1/A9 cells characterized by reversible MHC­I downregulation. However, the composition of the cell infiltrate was different and, most importantly, infiltration with immune cells was not increased in TC­1/dB2m tumors after immunotherapy. Therefore, the TC­1/dB2m cell line represents a clinically relevant tumor model that may be used for enhancement of cancer immunotherapy.


Assuntos
Linhagem Celular Tumoral/metabolismo , Antígenos de Histocompatibilidade Classe I/imunologia , Neoplasias/imunologia , Animais , Regulação Neoplásica da Expressão Gênica/genética , Antígenos de Histocompatibilidade Classe I/genética , Humanos , Imunoterapia , Interferon gama/imunologia , Camundongos , Neoplasias/genética , Neoplasias/patologia , Linfócitos T Citotóxicos/imunologia , Linfócitos T Citotóxicos/patologia , Microglobulina beta-2/genética , Microglobulina beta-2/imunologia
20.
Mol Cell ; 76(5): 838-851.e5, 2019 12 05.
Artigo em Inglês | MEDLINE | ID: mdl-31564558

RESUMO

Intermediary metabolism in cancer cells is regulated by diverse cell-autonomous processes, including signal transduction and gene expression patterns, arising from specific oncogenotypes and cell lineages. Although it is well established that metabolic reprogramming is a hallmark of cancer, we lack a full view of the diversity of metabolic programs in cancer cells and an unbiased assessment of the associations between metabolic pathway preferences and other cell-autonomous processes. Here, we quantified metabolic features, mostly from the 13C enrichment of molecules from central carbon metabolism, in over 80 non-small cell lung cancer (NSCLC) cell lines cultured under identical conditions. Because these cell lines were extensively annotated for oncogenotype, gene expression, protein expression, and therapeutic sensitivity, the resulting database enables the user to uncover new relationships between metabolism and these orthogonal processes.


Assuntos
Carcinoma Pulmonar de Células não Pequenas/metabolismo , Carcinoma Pulmonar de Células não Pequenas/patologia , Linhagem Celular Tumoral/metabolismo , Metaboloma/fisiologia , Biomarcadores Tumorais/metabolismo , Cromatografia Gasosa-Espectrometria de Massas/métodos , Regulação Neoplásica da Expressão Gênica/fisiologia , Glucose/metabolismo , Glutamina/metabolismo , Humanos , Redes e Vias Metabólicas/genética , Metabolômica/métodos , Neoplasias/metabolismo
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