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1.
Oncol Rep ; 42(5): 2029-2038, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31432145

RESUMO

In vitro culture of patient­derived tumor cells offers many advantages in the development of novel therapies for colorectal cancer. Although various culture systems have been developed, the long­term expansion of patient­derived tumor cells remains challenging. The present results suggested that tumor cells isolated from colorectal cancer patient­derived xenografts can be efficiently immortalized in conditioned medium from irradiated feeder cells containing Y­27632, a rho­associated coiled­coil containing protein kinase (ROCK) inhibitor. Patient­derived tumor cells proliferated rapidly, reaching 90­95% confluence in ~6 days. Short tandem repeat analysis suggested that these tumor tissues and cultured cells presented 13 identical short tandem repeat loci, including Amelogenin, Penta E, Penta D, D2S1338 and D19S433. Their epithelial phenotype was confirmed by staining for epithelial cell adhesion molecule and cytokeratin 20, whereas vimentin was used as a mesenchymal marker. When cells were transferred to 3D cultures, they continued to proliferate, forming well­defined tumor spheroids. Expression levels of human telomerase reverse transcriptase and C­Myc mRNA were increased in cultured cells. Finally, immortalized cells were used for the screening of 65 anticancer drugs approved by the Food and Drug Administration, allowing the identification of gene­drug associations. In the present study, primary culture models of colorectal cancer were efficiently established using a ROCK inhibitor and feeder cells, and this approach could be used for personalized treatment strategies for patients with colorectal cancer.


Assuntos
Amidas/farmacologia , Neoplasias Colorretais/patologia , Células Alimentadoras/citologia , Cultura Primária de Células/métodos , Piridinas/farmacologia , Células Tumorais Cultivadas/citologia , Idoso , Animais , Proliferação de Células , Neoplasias Colorretais/genética , Neoplasias Colorretais/metabolismo , Meios de Cultivo Condicionados/química , Feminino , Humanos , Masculino , Camundongos , Repetições de Microssatélites , Pessoa de Meia-Idade , Transplante de Neoplasias , Esferoides Celulares/citologia , Esferoides Celulares/metabolismo , Células Tumorais Cultivadas/efeitos dos fármacos
2.
Electron. j. biotechnol ; 40: 58-64, July. 2019. graf, tab, ilus
Artigo em Inglês | LILACS | ID: biblio-1053475

RESUMO

Background: Prodigiosin has been demonstrated to be an important candidate in investigating anticancer drugs and in many other applications in recent years. However, industrial production of prodigiosin has not been achieved. In this study, we found a prodigiosin-producing strain, Serratia marcescens FZSF02, and its fermentation strategies were studied to achieve the maximum yield of prodigiosin. Results: When the culture medium consisted of 16.97 g/L of peanut powder, 16.02 g/L of beef extract, and 11.29 mL/L of olive oil, prodigiosin reached a yield of 13.622 ± 236 mg/L after culturing at 26 °C for 72 h. Furthermore, when 10 mL/L olive oil was added to the fermentation broth at the 24th hour of fermentation, the maximum prodigiosin production of 15,420.9 mg/L was obtained, which was 9.3-fold higher than the initial level before medium optimization. More than 60% of the prodigiosin produced with this optimized fermentation strategy was in the form of pigment pellets. To the best of our knowledge, this is the first report on this phenomenon of pigment pellet formation, which made it much easier to extract prodigiosin at low cost. Prodigiosin was then purified and identified by absorption spectroscopy, HPLC, and LCMS. Purified prodigiosin obtained in this study showed anticancer activity in separate experiments on several human cell cultures: A549, K562, HL60, HepG2, and HCT116. Conclusions: This is a promising strain for producing prodigiosin. The prodigiosin has potential in anticancer medicine studies.


Assuntos
Prodigiosina/biossíntese , Prodigiosina/farmacologia , Serratia marcescens/metabolismo , Antineoplásicos/farmacologia , Arachis/química , Pós , Prodigiosina/isolamento & purificação , Espectrometria de Massas , Células Tumorais Cultivadas/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Técnicas de Cultura de Células , Fermentação , Azeite de Oliva/química , Acetatos , Nitrogênio
3.
Int Urol Nephrol ; 51(10): 1771-1779, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31236854

RESUMO

OBJECTIVES: Radiotherapy is the primary option for bladder cancer patients, but it does not have obvious curative effects. This study was to investigate how to increase radiosensitivity in bladder cancer. MATERIALS AND METHODS: The curcumin and irradiation treated T24 cells were used for analysis of microRNA expression (miRNA microarray), cell viability (Cell Proliferation Assay Kit), colony formation, apoptosis (Annexin V-FITC/7-AAD flow cytometry), miR-1246 and p53 mRNA (real-time PCR) and protein (Western blot) expression. RESULTS: Microarray assay identified 17 differentially expressed miRNAs (twofold change) in curcumin treated cells compared to control cells. Among them, miR-1246 was the miRNA with the largest change in expression after curcumin treatment. Curcumin significantly decreased T24 cell viability and colony formation in a concentration-dependent manner compared to control cells. miR-1246 expression was significantly higher in T24 cells than in SV-HUC-1 cells and the higher concentrations (10 or 20 µM) of curcumin significantly down-regulated miR-1246 expression in T24 and HT-1376 cells. The combination of 10 µM curcumin and irradiation was more effective in decreasing miR-1246 expression, cell viability and colony formation than curcumin or irradiation alone. Inhibition of miR-1246 significantly decreased cell viability and colony formation in T24 and HT-1376 cells. Transfection with antagomiR-1246 significantly increased the G0/G1-phase of T24 cells and induced apoptosis compared to cells transfected with antagomiR-NC. Luciferase reporter assay showed that the overexpression of miR-1246 suppressed the luciferase activity of the P53 3'-UTR reporter genes. CONCLUSION: miR-1246 is involved in the anti-cancer effects of curcumin and irradiation through targeting the inhibition of p53 gene translation in bladder cancer cells.


Assuntos
Antineoplásicos/farmacologia , Curcumina/farmacologia , MicroRNAs/fisiologia , Radiossensibilizantes/farmacologia , Proteína Supressora de Tumor p53/efeitos dos fármacos , Proteína Supressora de Tumor p53/fisiologia , Neoplasias da Bexiga Urinária/patologia , Humanos , Células Tumorais Cultivadas/efeitos dos fármacos , Neoplasias da Bexiga Urinária/radioterapia
4.
BMC Cancer ; 19(1): 628, 2019 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-31238897

RESUMO

BACKGROUND: A major barrier to effective treatment of glioblastoma (GBM) is the large intertumoral heterogeneity at the genetic and cellular level. In early phase clinical trials, patient heterogeneity in response to therapy is commonly observed; however, how tumor heterogeneity is reflected in individual drug sensitivities in the treatment-naïve glioblastoma stem cells (GSC) is unclear. METHODS: We cultured 12 patient-derived primary GBMs as tumorspheres and validated tumor stem cell properties by functional assays. Using automated high-throughput screening (HTS), we evaluated sensitivity to 461 anticancer drugs in a collection covering most FDA-approved anticancer drugs and investigational compounds with a broad range of molecular targets. Statistical analyses were performed using one-way ANOVA and Spearman correlation. RESULTS: Although tumor stem cell properties were confirmed in GSC cultures, their in vitro and in vivo morphology and behavior displayed considerable tumor-to-tumor variability. Drug screening revealed significant differences in the sensitivity to anticancer drugs (p < 0.0001). The patient-specific vulnerabilities to anticancer drugs displayed a heterogeneous pattern. They represented a variety of mechanistic drug classes, including apoptotic modulators, conventional chemotherapies, and inhibitors of histone deacetylases, heat shock proteins, proteasomes and different kinases. However, the individual GSC cultures displayed high biological consistency in drug sensitivity patterns within a class of drugs. An independent laboratory confirmed individual drug responses. CONCLUSIONS: This study demonstrates that patient-derived and treatment-naïve GSC cultures maintain patient-specific traits and display intertumoral heterogeneity in drug sensitivity to anticancer drugs. The heterogeneity in patient-specific drug responses highlights the difficulty in applying targeted treatment strategies at the population level to GBM patients. However, HTS can be applied to uncover patient-specific drug sensitivities for functional precision medicine.


Assuntos
Neoplasias Encefálicas/tratamento farmacológico , Glioblastoma/tratamento farmacológico , Ensaios de Triagem em Larga Escala , Células-Tronco Neoplásicas/efeitos dos fármacos , Esferoides Celulares/efeitos dos fármacos , Células Tumorais Cultivadas/efeitos dos fármacos , Animais , Antineoplásicos/farmacologia , Neoplasias Encefálicas/diagnóstico por imagem , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/patologia , Ensaios de Seleção de Medicamentos Antitumorais/métodos , Feminino , Glioblastoma/diagnóstico por imagem , Glioblastoma/genética , Glioblastoma/patologia , Humanos , Camundongos SCID , Transplante de Neoplasias , Células-Tronco Neoplásicas/patologia , Esferoides Celulares/patologia , Células Tumorais Cultivadas/patologia
5.
Acta Biomed ; 90(2): 241-247, 2019 05 23.
Artigo em Inglês | MEDLINE | ID: mdl-31125002

RESUMO

BACKGROUND AND AIM OF THE STUDY: Use of commercial products containing nanoparticles formulated from zinc oxide (ZnO) and aluminium oxide (Al2O-3) has increased significantly. These nanoparticles are widely used as ingredient in cosmetics, and also in food packaging industry although their toxicity status is yet to be studied. Here, we aimed to explore the effect of zinc oxide nanoparticles (ZnO-NPs) and aluminium oxide nanoparticles (ANPs) in human HT29 colon cancer cell line. METHODS: In this study, ZnO-NPs were synthesized by chemical method and ANPs synthesized by sol-gel method and were characterized using UV-Vis spectroscopy, X ray diffraction and Transmittance electron microscopy. The effects of ZnO-NPs and ANPs was determined by cell viability, membrane integrity and colony formation potentials. RESULTS: ZnO-NPs and ANPs inhibit HT29, colon cancer cell proliferation in a dose dependent manner, and affect the membrane potentials and also prevent the colony formation. CONCLUSIONS: The results suggest that ZnO NPs are found to be more effective than ANPs in reducing colon cancer cell proliferation.


Assuntos
Óxido de Alumínio/farmacologia , Nanopartículas/administração & dosagem , Óxido de Zinco/farmacologia , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Neoplasias do Colo/tratamento farmacológico , Neoplasias do Colo/patologia , Humanos , Sensibilidade e Especificidade , Células Tumorais Cultivadas/efeitos dos fármacos
6.
Cancer Med ; 8(4): 1793-1805, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30843650

RESUMO

Antibody-drug conjugates (ADCs) belong to a promising class of biopharmaceuticals in which target-killing of tumor cells was achieved by marrying the potency of the cytotoxic payload with the tumor specificity of the antibody. Here we developed a novel ADC (ZV0508) that targets 5T4 oncofetal antigen, which is overexpressed in many carcinomas on both bulk tumor cells and cancer stem cells. A novel cytotoxic payload called Duostatin-5 (Duo-5) which was derived from monomethyl auristatin F (MMAF) was attached to a 5T4 targeting antibody (ZV05) by interchain cysteine cross-linking conjugation via a disubstituted C-Lock linker. We have investigated the antitumor efficacy of ZV0508 by in vitro and in vivo studies, and compared its antitumor activity with ZV05-mcMMAF (ZV0501), in which MMAF was linked via a conventional noncleavable maleimidocaproyl linker. As results, ZV0508 exhibited ideal antiproliferative effects through blocking cell cycle and inducing cell apoptosis. The in vivo studies revealed that both ZV0501 and ZV0508 exhibited excellent antitumor activities even at a single dose. Although ZV0508 was inferior to ZV0501 in vitro, it elicited more durable antitumor responses than ZV0501 in vivo. The superior in vivo activity of ZV0508 may be due to the combined use of the disubstituted C-Lock linker and the novel payload Duo-5, resulting in a more stable and potent ADC. Taken together, these data suggest ZV0508 is a worthy candidate for the treatment of 5T4 positive cancers.


Assuntos
Antígenos de Neoplasias/metabolismo , Antineoplásicos/farmacologia , Imunoconjugados/farmacologia , Neoplasias/patologia , Animais , Anticorpos Monoclonais/metabolismo , Anticorpos Monoclonais/farmacologia , Anticorpos Monoclonais/uso terapêutico , Antineoplásicos/farmacocinética , Antineoplásicos/uso terapêutico , Apoptose/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Reagentes para Ligações Cruzadas , Feminino , Humanos , Imunoconjugados/farmacocinética , Imunoconjugados/uso terapêutico , Masculino , Neoplasias Mamárias Experimentais/tratamento farmacológico , Neoplasias Mamárias Experimentais/metabolismo , Neoplasias Mamárias Experimentais/patologia , Camundongos Endogâmicos BALB C , Camundongos Nus , Terapia de Alvo Molecular/métodos , Neoplasias/tratamento farmacológico , Neoplasias/metabolismo , Oligopeptídeos/farmacologia , Células Tumorais Cultivadas/efeitos dos fármacos , Ensaios Antitumorais Modelo de Xenoenxerto
7.
J Coll Physicians Surg Pak ; 29(3): 240-244, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30823950

RESUMO

OBJECTIVE: To investigate the expression of zinc finger transcription factors-Snail and E-cadherin in adriamycin-resistant human breast cancer MCF-7/ADM cells and non-resistant MCF-7 cells. STUDY DESIGN: An experimental study. PLACE AND DURATION OF STUDY: The Affiliated Cancer Hospital of Nanjing Medical University and Jiangsu Cancer Hospital, and Jiangsu Institute of Cancer Research, China, from April 2017 to March 2018. METHODOLOGY: Real-time quantitative PCR technology was used to detect the expression levels of Snail mRNA and E-cadherin mRNA in normal breast cells, adriamycin-resistant human breast cancer MCF-7/ADM cells and non-resistant MCF-7 cells. Western blot was used to detect the expression levels of proteins of Snail and E-cadherin in normal breast cells, adriamycin-resistant human breast cancer MCF-7/ADM cells and non-resistant MCF-7 cells. RESULTS: The expression of Snail mRNA and protein in adriamycin-resistant human breast cancer MCF-7/ADM cells was significantly higher than that in normal breast cells (p<0.001) and non-resistant MCF-7 cells (p<0.001). The expression of E-cadherin mRNA and protein in adriamycin-resistant human breast cancer MCF-7/ADM cells was significantly lower than that in normal breast cells (p<0.001) and non-resistant MCF-7 cells (p<0.001). CONCLUSION: Adriamycin-resistant human breast cancer MCF-7/ADM cell strains had high expression of Snail and low expression of E-cadherin. This points out to a new research direction for the targeted therapy of drug-resistant breast cancer cells, and provides clinical guidance for breast cancer therapy and prognosis evaluation.


Assuntos
Neoplasias da Mama/genética , Caderinas/genética , Doxorrubicina/farmacologia , Fatores de Transcrição da Família Snail/genética , Western Blotting , Neoplasias da Mama/patologia , China , Resistencia a Medicamentos Antineoplásicos , Feminino , Regulação Neoplásica da Expressão Gênica , Hospitais Universitários , Humanos , Células MCF-7/efeitos dos fármacos , Células MCF-7/metabolismo , Terapia de Alvo Molecular , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Valores de Referência , Células Tumorais Cultivadas/efeitos dos fármacos
8.
J Ovarian Res ; 12(1): 15, 2019 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-30736825

RESUMO

BACKGROUND: Ovarian cancer is the most lethal of all gynecologic malignancies. The relationship between sexual steroids receptors and ovarian cancer progression has been largely evaluated. The presence of progesterone receptors has been associated with an increase of a disease-free period and overall survival in patients with ovarian carcinoma. In the present study, primary cultures of ovarian carcinoma obtained from 35 patients diagnosed with epithelial ovarian cancer were evaluated for cell survival after treatment with 10- 8 M of 17ß-estradiol, progesterone, testosterone and dihydrotestosterone. RESULTS: The results were analyzed considering histological subtypes: low grade serous, high grade serous, endometrioid and mucinous carcinoma; clear cell carcinoma was not included due to failure in obtaining successful cultures of this subtype. A significant reduction of cell survival was observed after progesterone treatment in endometrioid ovarian carcinoma. Changes were not observed in low grade serous, high grade serous and mucinous carcinoma. The effect of progesterone was related to the presence of progesterone receptor (PR), a 43% reduction in the cell number was observed in PR (+) endometrioid ovarian carcinoma. CONCLUSIONS: This study supports the importance of progesterone and the presence of progesterone receptor in the reduction of ovarian cancer progression in the endometrioid ovarian carcinoma.


Assuntos
Antineoplásicos Hormonais/farmacologia , Carcinoma Endometrioide/patologia , Sobrevivência Celular/efeitos dos fármacos , Neoplasias Ovarianas/patologia , Progesterona/farmacologia , Adulto , Carcinoma Endometrioide/metabolismo , Carcinoma Epitelial do Ovário/metabolismo , Carcinoma Epitelial do Ovário/patologia , Proliferação de Células/efeitos dos fármacos , Feminino , Humanos , Pessoa de Meia-Idade , Neoplasias Ovarianas/metabolismo , Estudos Prospectivos , Receptores de Progesterona/metabolismo , Receptores de Esteroides/metabolismo , Células Tumorais Cultivadas/efeitos dos fármacos
9.
J Control Release ; 295: 21-30, 2019 02 10.
Artigo em Inglês | MEDLINE | ID: mdl-30550941

RESUMO

Diffuse large B cell lymphoma (DLBCL), the most common subtype of Non-Hodgkin lymphoma, exhibits pathologic heterogeneity and a dynamic immunogenic tumor microenvironment (TME). However, the lack of preclinical in vitro models of DLBCL TME hinders optimal therapeutic screening. This study describes the development of an integrated droplet microfluidics-based platform for high-throughput generation of immunogenic DLBCL spheroids. The spheroids consist of three cell types (cancer, fibroblast and lymphocytes) in a novel hydrogel combination of alginate and puramatrix, which promoted cell adhesion and aggregation. This system facilitates dynamic analysis of cellular interaction, proliferation and therapeutic efficacy via spatiotemporal monitoring and secretome profiling. The immunomodulatory drug lenalidomide had direct anti-proliferative effect on activated B-cell like DLBCL spheroids and reduced several cytokines and other markers (e.g., CCL2, CCL3, CCL4, CD137 and ANG-1 levels) compared with untreated spheroids. Collectively, this novel spheroid platform will enable high-throughput anti-cancer therapeutic screening in a semi-automated manner.


Assuntos
Técnicas de Cultura de Células/métodos , Ensaios de Seleção de Medicamentos Antitumorais/métodos , Dispositivos Lab-On-A-Chip , Linfoma Difuso de Grandes Células B/tratamento farmacológico , Esferoides Celulares/efeitos dos fármacos , Alginatos/química , Antineoplásicos/farmacologia , Biomarcadores Tumorais/análise , Biomarcadores Tumorais/imunologia , Técnicas de Cultura de Células/instrumentação , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Técnicas de Cocultura/instrumentação , Técnicas de Cocultura/métodos , Ensaios de Seleção de Medicamentos Antitumorais/instrumentação , Desenho de Equipamento , Humanos , Hidrogéis/química , Fatores Imunológicos/farmacologia , Lenalidomida/farmacologia , Linfoma Difuso de Grandes Células B/imunologia , Esferoides Celulares/imunologia , Células Tumorais Cultivadas/efeitos dos fármacos , Células Tumorais Cultivadas/imunologia
10.
J Appl Toxicol ; 39(1): 38-71, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30073673

RESUMO

Development of a cancer is a multistep process and six major hallmarks of cancer that are known to control malignant transformation have been described. Anticancer drug development is a tedious process, requiring a number of in vitro, in vivo and clinical studies. In vitro assays provide an initial platform for cancer drug discovery approaches. A wide range of in vitro assays/techniques have been developed to evaluate each hallmark feature of cancer and selection of a particular in vitro assay or technique mainly depends on the specific research question (s) to be examined. In the present review, we have described some commonly utilized in vitro assays and techniques used to examine cell viability/proliferation, apoptosis, cellular senescence, invasion and migration, oxidative stress and antioxidant effects, gene and protein expression, angiogenesis and genomic alterations in cancer drug discovery. Additionally, uses of modern techniques such as high throughput screening, high content screening and reporter gene assays in cancer drug discovery have also been described.


Assuntos
Antineoplásicos/farmacologia , Proliferação de Células/efeitos dos fármacos , Descoberta de Drogas/métodos , Avaliação Pré-Clínica de Medicamentos/métodos , Neoplasias/tratamento farmacológico , Células Tumorais Cultivadas/efeitos dos fármacos , Humanos
11.
Mem Inst Oswaldo Cruz ; 113(11): e180267, 2018 Oct 11.
Artigo em Inglês | MEDLINE | ID: mdl-30328891

RESUMO

The Bacille Calmette-Guérin (BCG) vaccine comprises a family of genetically different strains derived by the loss of genomic regions (RDs) and other mutations. In BCG Moreau, loss of RD16 inactivates rv3405c * , encoding a transcriptional repressor that negatively regulates the expression of Rv3406, an alkyl sulfatase. To evaluate the impact of this loss on the BCG and host cell viability and the cytokine profile, THP-1 cells were infected with BCG Moreau (harbouring the empty vector) and a complemented strain carrying a functional copy of rv3405c. Viability of the host cells and bacteria as well as the pattern of cytokine secretion were evaluated. Our results show that the viability of BCG Moreau is higher than that of the complemented strain in an axenic medium, suggesting a possible functional gain associated with the constitutive expression of Rv3406. Viability of the host cells did not vary significantly between recombinant strains, but differences in the profiles of the cytokine secretion (IL-1ß and IL-6) were observed. Our results suggest an example of a functional gain due to gene loss contributing to the elucidation of the impact of RD16 on the physiology of BCG Moreau.


Assuntos
Vacina BCG/farmacologia , Sobrevivência Celular/genética , Citocinas/efeitos dos fármacos , Mutação com Ganho de Função/genética , Macrófagos/microbiologia , Mycobacterium bovis/genética , Transcrição Genética/genética , Vacina BCG/genética , Sobrevivência Celular/efeitos dos fármacos , Citocinas/genética , Mutação com Ganho de Função/efeitos dos fármacos , Humanos , Mycobacterium bovis/fisiologia , Fatores de Tempo , Transcrição Genética/efeitos dos fármacos , Células Tumorais Cultivadas/efeitos dos fármacos , Células Tumorais Cultivadas/microbiologia
12.
Br J Haematol ; 183(2): 196-211, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-30080238

RESUMO

CD38 is expressed on Waldenström macroglobulinaemia (WM) cells, but its role as a therapeutic target remains undefined. With recent approval of the anti-CD38 monoclonal antibody, daratumumab (Dara), we hypothesized that blocking CD38 would be lethal to WM cells. In vitro Dara treatment of WM cells (including ibrutinib-resistant lines) elicited antibody-dependent cellular cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC), antibody-dependent cell phagocytosis (ADCP) and direct apoptosis. In vivo, Dara treatment was well tolerated and delayed tumour growth in RPCI-WM1-xenografted mice. CD38 is reported to augment B-cell receptor (BCR) signalling; we noted that Dara significantly attenuated phosphorylated SYK, LYN, BTK, PLCγ2, ERK1/2, AKT, mTOR, and S6 levels, and this effect was augmented by cotreatment with ibrutinib. Indeed, WM cells, including ibrutinib-resistant WM cell lines treated with the ibrutinib + Dara combination, showed significantly more cell death through ADCC, CDC, ADCP and apoptosis relative to single-agent Dara or ibrutinib. In summary, we are the first to report the in vitro and in vivo anti-WM activity of Dara. Furthermore, we show a close connection between BCR and CD38 signalling, which can be co-targeted with ibrutinib + Dara to induce marked WM cell death, irrespective of acquired resistance to ibrutinib.


Assuntos
ADP-Ribosil Ciclase 1/antagonistas & inibidores , Anticorpos Monoclonais/farmacologia , Antineoplásicos/farmacologia , Macroglobulinemia de Waldenstrom/patologia , Animais , Anticorpos Monoclonais/administração & dosagem , Anticorpos Monoclonais/uso terapêutico , Antineoplásicos/uso terapêutico , Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Citotoxicidade Imunológica/efeitos dos fármacos , Humanos , Camundongos Endogâmicos NOD , Fagocitose/efeitos dos fármacos , Pirazóis/administração & dosagem , Pirimidinas/administração & dosagem , Células Tumorais Cultivadas/efeitos dos fármacos , Macroglobulinemia de Waldenstrom/imunologia , Macroglobulinemia de Waldenstrom/prevenção & controle , Ensaios Antitumorais Modelo de Xenoenxerto
13.
Clin Neurol Neurosurg ; 173: 20-30, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-30055402

RESUMO

OBJECTIVE: Glial tumor growth may accelerate during gestation, but epidemiological studies consistently demonstrated that parousity reduces life long risk of glial tumors. Pregnancy may also accelerate growth of medulloblastoma and meningioma, but parousity does not confer protection against these tumors. We were the first to show that medroxyprogesterone acetate (MPA) reduces rat C6 glioma growth in vitro. Now we aimed to determine the effects of MPA on human brain cancers (particularly glioblastoma) in vitro and C6 glioma in vivo. PATIENTS AND METHODS: We evaluated the effects of MPA on: i) monolayer growth of human U87 and U251 glioblastoma, ii) 3D-spheroid growth and invasion of C6 rat glioma and human U251 glioma, iii) interactions with PI3-Kinase inhibitors and coxsackie-adenovirus receptor (CAR) in modifying 3D-spheroid invasion of glioma. RESULTS: MPA at low doses (3.25-13 µM) insignificantly stimulated and at high doses (above 52 µM) strongly suppressed the growth of human U87 and U251 cells in vitro. MPA also binds to glucocorticoid receptors similar to dexamethasone (Dex) and unexpectedly, PI3-Kinase inhibitors at low doses suppressed anti-invasive efficacies of MPA and Dex. MPA exerted higher invasion-inhibitory effects on CAR-expressing human glioma cells. Lastly, MPA suppressed growth of C6 glioma implanted into rat brain. CONCLUSION: Progesterone analogues deserve to be studied in future experimental models of high grade glial brain tumors.


Assuntos
Neoplasias Encefálicas/tratamento farmacológico , Glioblastoma/tratamento farmacológico , Glioma/tratamento farmacológico , Medroxiprogesterona/farmacologia , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/patologia , Neoplasias Encefálicas/patologia , Neoplasias Cerebelares/tratamento farmacológico , Dexametasona/farmacologia , Modelos Animais de Doenças , Glioblastoma/patologia , Glioma/patologia , Humanos , Meningioma/tratamento farmacológico , Ratos , Células Tumorais Cultivadas/efeitos dos fármacos
14.
Analyst ; 143(14): 3317-3326, 2018 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-29931010

RESUMO

In the present study, human primary oral squamous carcinoma cells treated with cisplatin and 5-fluorouracil were analyzed, for the first time, by in vitro FTIR Microspectroscopy (FTIRM), to improve the knowledge on the biochemical pathways activated by these two chemotherapy drugs. To date, most of the studies regarding FTIRM cellular analysis have been executed on fixed cells from immortalized cell lines. FTIRM analysis performed on primary tumor cells under controlled hydrated conditions provides more reliable information on the biochemical processes occurring in in vivo tumor cells. This spectroscopic analysis allows to get on the same sample and at the same time an overview of the composition and structure of the most remarkable cellular components. In vitro FTIRM analysis of primary oral squamous carcinoma cells evidenced a time-dependent drug-specific cellular response, also including apoptosis triggering. Furthermore, the univariate and multivariate analyses of IR data evidenced meaningful spectroscopic differences ascribable to alterations affecting cellular proteins, lipids and nucleic acids. These findings suggest for the two drugs different pathways and extents of cellular damage, not provided by conventional cell-based assays (MTT assay and image-based cytometry).


Assuntos
Antineoplásicos/farmacologia , Carcinoma de Células Escamosas/patologia , Cisplatino/farmacologia , Fluoruracila/farmacologia , Espectroscopia de Infravermelho com Transformada de Fourier , Apoptose , Humanos , Células Tumorais Cultivadas/efeitos dos fármacos
15.
Oncology ; 94(5): 324-328, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29533961

RESUMO

The metallo-phosphorus dendrimer 1G3-Cu (generation 3 dendrimer bearing 48 conjugated copper(II) on its surface) has antiproliferative activity related to its capacity to activate Bax translocation. In the present study, we evaluate the activity of an association of 1G3-Cu with 5 cytotoxic agents used in chemotherapy having different modes of action. Data show no additive effect with camptothecin and cisplatin, additivity with paclitaxel and MG132, and synergy with doxorubicin. Results suggest that the multivalent Cu-conjugated dendrimer 1G3-Cu (activator of Bax translocation) plays an important role in boosting the clinical impact of Bax accumulation stimulated by the proteasome inhibitor MG132, antimitotic taxanes, and the topo II inhibitor doxorubicin.


Assuntos
Antineoplásicos/farmacologia , Transporte Biológico Ativo/efeitos dos fármacos , Linhagem Celular Tumoral/efeitos dos fármacos , Citotoxinas/farmacologia , Células Tumorais Cultivadas/efeitos dos fármacos , Antineoplásicos/síntese química , Camptotecina , Proliferação de Células , Cisplatino , Cobre/farmacologia , Citotoxinas/síntese química , Doxorrubicina , Humanos , Nanomedicina , Paclitaxel , Fósforo/farmacologia , Taxoides
16.
Tissue Cell ; 50: 15-30, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29429514

RESUMO

Alternative models such as three-dimensional (3D) cell cultures represent a distinct milestone towards capturing the realities of cancer biology in vitro and reduce animal experimentation in the preclinical stage of drug discovery. Significant work remains to be done to understand how substrates used in in vitro alternatives influence cancer cells phenotype and drug efficacy responses, so that to accurately link such models to specific in vivo disease scenarios. Our study describes how the morphological, mechanical and biochemical properties of adenocarcinoma (A549) cells change in response to a 3D environment and varying substrates. Confocal Laser Scanning (LSCM), He-Ion (HIM) and Atomic Force (AFM) microscopies, supported by ELISA and Western blotting, were used. These techniques enabled us to evaluate the shape, cytoskeletal organization, roughness, stiffness and biochemical signatures of cells grown within soft 3D matrices (PuraMatrix™ and Matrigel™), and to compare them to those of cells cultured on two-dimensional glass substrates. Cell cultures are also characterized for their biological response to docetaxel, a taxane-type drug used in Non-Small-Cell Lung Cancer (NSCLC) treatment. Our results offer an advanced biophysical insight into the properties and potential application of 3D cultures of A549 cells as in vitro alternatives in lung cancer research.


Assuntos
Adenocarcinoma/tratamento farmacológico , Fenômenos Biofísicos , Técnicas de Cultura de Células/métodos , Neoplasias Pulmonares/tratamento farmacológico , Células Tumorais Cultivadas/ultraestrutura , Células A549 , Adenocarcinoma/química , Adenocarcinoma/patologia , Adenocarcinoma de Pulmão , Docetaxel , Ensaio de Imunoadsorção Enzimática , Humanos , Neoplasias Pulmonares/química , Neoplasias Pulmonares/patologia , Microscopia Confocal , Especificidade por Substrato , Taxoides/farmacologia , Células Tumorais Cultivadas/química , Células Tumorais Cultivadas/efeitos dos fármacos
17.
Dent Mater ; 34(3): 519-530, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29373133

RESUMO

OBJECTIVES: Camphorquinone (CQ) is the most important photoinitiator used in dental composite resins. Sparse data indicate a mutagenic potential of CQ. Therefore, it was aim of this study to evaluate the cytotoxicity, genotoxicity, and mutagenicity of CQ in L5178Y TK+/- mouse lymphoma cells. METHODS: L5178Y/TK+/- cells were exposed to different concentrations of non-irradiated CQ (0.25-2.5mM). Cytotoxicity was evaluated by propidium iodide assay, determination of suspension growth rate, relative total growth and the mitotic index. Intracellular levels of reactive oxygen/nitrogen species (ROS/RNS) were quantified by 2',7'-dichlorofluoresceine diacetate (DCFH-DA). Early induction of DNA strand breaks and oxidative DNA base lesions was assessed using the 8-hydroxyguanine DNA-glycosylase 1 (hOGG1)-modified alkaline comet assay, whereas mutagenicity of CQ was determined in the mouse lymphoma TK assay (MLA), according to OECD Guideline No. 490. RESULTS: CQ (0.5-2.5mM) induced concentration- and time-dependent inhibition of cell growth associated with increased ROS/RNS production, amounting to 2342%±1108% of controls after 90min at 2.5mM. Additionally, CQ concentration-dependently caused direct DNA-damage, i.e. formation of DNA strand breaks and 8-hydroxy-2'-deoxyguanosine. Whereas the MLA indicated lack of mutagenicity of CQ after a 4h of treatment, CQ concentration-dependently increased total mutant frequency (MF) after 24h (about 2-fold at 2.5mM). But, based on the global evaluation factor concept, increase in MF did not reach biologically relevance. SIGNIFICANCE: CQ induced concentration-dependent, cytotoxic and genotoxic effects in L5178Y/TK+/- cells, most likely due to oxidative stress, but without mediating obvious biological relevant mutagenicity.


Assuntos
Cânfora/análogos & derivados , Células Tumorais Cultivadas/efeitos dos fármacos , Animais , Cânfora/toxicidade , Sobrevivência Celular/efeitos dos fármacos , Ensaio Cometa , Dano ao DNA/efeitos dos fármacos , Técnicas In Vitro , Linfoma , Camundongos , Índice Mitótico , Testes de Mutagenicidade , Estresse Oxidativo , Espécies Reativas de Oxigênio/metabolismo
18.
J Clin Lab Anal ; 32(5): e22379, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29333615

RESUMO

BACKGROUND: Recent studies have revealed that circular RNAs are involved in the biological process of some kinds of human cancers. However, little is known about their diagnostic values and functions in colorectal cancer (CRC). METHODS: The expression levels of hsa_circ_0000567 in 102 paired CRC tissues and adjacent noncancerous tissues, 5 CRC cell lines, and a normal colorectal epithelial cell line were detected by quantitative real-time polymerase chain reaction (qRT-PCR). The correlations between hsa_circ_0000567 expression levels and the clinicopathological factors of patients with CRC were analyzed. Furthermore, the loss-of-function assay was performed to investigate the functions of hsa_circ_0000567 in vitro. Finally, a receiver operating characteristic (ROC) curve was established to evaluate the diagnostic value of hsa_circ_0000567. RESULTS: Hsa_circ_0000567 expression was significantly downregulated in CRC tissues and CRC cell lines. In addition, the decreased hsa_circ_0000567 expression in CRC was negatively correlated with tumor size (P = .011), lymph metastasis (P = .003), distal metastasis (P < .0001), and tumor-node-metastasis (TNM) stage (P = .003) in CRC. Moreover, knockdown of hsa_circ_0000567 promoted CRC cells proliferation and migration in vitro. Importantly, the area under the ROC curve (AUC) was 0.8653, which indicates hsa_circ_0000567 can serve as a diagnostic biomarker. CONCLUSION: Hsa_circ_0000567 may be a novel suppressor and a potential diagnosis biomarker in CRC.


Assuntos
Biomarcadores Tumorais/metabolismo , Neoplasias Colorretais/diagnóstico , Regulação Neoplásica da Expressão Gênica/fisiologia , RNA/genética , Adulto , Fatores Etários , Idoso , Antibióticos Antineoplásicos/farmacologia , Movimento Celular/efeitos dos fármacos , Proliferação de Células/fisiologia , Análise Mutacional de DNA , Dactinomicina/farmacologia , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Masculino , Pessoa de Meia-Idade , RNA Circular , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Curva ROC , Transfecção , Células Tumorais Cultivadas/efeitos dos fármacos
19.
Cancer Chemother Pharmacol ; 81(3): 469-481, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29308536

RESUMO

PURPOSE: Esophageal squamous cell carcinoma (ESCC) is one of the most aggressive squamous cell carcinomas and is generally resistant to chemotherapy. In the present study, the cytotoxic activity of Rabdocoestin B (Rabd-B) against ESCC and the underlying mechanisms were investigated. METHODS: The inhibitory effect of Rabd-B on KYSE30 and KYSE450 was evaluated by Cell Counting Kit-8 (CCK8) and colony formation assays in vitro. The cell cycle distribution and apoptosis of cells treated with Rabd-B were determined by flow cytometry. The mechanisms underlying the effects of Rabd-B were systematically examined by Western blot. The in vivo anti-tumor ability of Rabd-B was measured in mouse xenograft models and cisplatin (DDP) was used as positive control. RESULTS: Rabd-B efficiently induced G2/M phase arrest in ESCC cells by upregulating the Chk1/Chk2-Cdc25C axis to inhibit the G2→M transition facilitated by Cdc2/Cyclin B1. Furthermore, Rabd-B suppressed ATM/ATR phosphorylation, thereby inhibiting BRCA1-mediated DNA repair, which resulted in mitotic catastrophe and induced cell apoptosis. Rabd-B also decreased the activity of the Akt and NF-κB survival signaling pathways and ultimately initiated the caspase-9-dependent intrinsic apoptotic pathway in ESCC cells. The apoptosis induced by Rabd-B could be partially reversed by a caspase-9-specific inhibitor (Z-LEHD-FMK) and a pan-caspase inhibitor (Z-VAD-FMK). Moreover, Rabd-B effectively suppressed tumor growth in mouse xenografts which was comparable to that of DDP without significant injuries to the mice. CONCLUSION: Taken together, these findings indicate that Rabd-B is a promising precursor compound that may be useful as a treatment for ESCC and thus warrants further investigation.


Assuntos
Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Diterpenos/farmacologia , Carcinoma de Células Escamosas do Esôfago/tratamento farmacológico , Animais , Antineoplásicos Fitogênicos/farmacologia , Proliferação de Células/efeitos dos fármacos , Neoplasias Esofágicas/tratamento farmacológico , Camundongos , Células Tumorais Cultivadas/efeitos dos fármacos , Células Tumorais Cultivadas/fisiologia , Ensaio Tumoral de Célula-Tronco/métodos , Ensaios Antitumorais Modelo de Xenoenxerto/métodos
20.
Rev. bras. cancerol ; 64(1): 93-98, Jan/Fev/Mar 2018.
Artigo em Português | LILACS | ID: biblio-969213

RESUMO

Introdução: Bisfosfonatos são fármacos utilizados para o tratamento de enfermidades ósseas, como a osteoporose e metástases ósseas, em razão do seu mecanismo de ação, que consiste na diminuição do processo de reabsorção do osso. Outros estudos verificaram que bisfosfonatos de alta potência, como o zoledronato, poderiam auxiliar no tratamento de outras enfermidades malignas por causa da promoção de um efeito antiproliferativo. Objetivo: Este estudo in vitro objetivou avaliar a atividade antiproliferativa de zoledronato em diferentes linhagens de células tumorais. Método: Nove linhagens humanas (U251; MCF7; NCI/ADR-RES; 786-0; NCI-H460; PC-3; OVCAR-3; HT29; K-562 e HaCaT) foram submetidas ao tratamento com as concentrações de 0,12; 1,2; 12 e 120 µM de zoledronato e tiveram sua atividade proliferativa avaliada após 48 horas, utilizando-se o corante sulforrodamina B. Resultados: Verificou-se que as concentrações de 12 µM e 120 µM de zoledronato foram eficazes para a redução em 50% e 100%, respectivamente, da proliferação das células 786-0 (carcinoma renal). A maior concentração de zoledronato (120 µM) promoveu um efeito citostático (redução da proliferação celular em 50%) para as células HaCaT (queratinócito humano não tumoral), HT-29 (carcinoma de cólon), NCI-ADR/ RES (adenocarcinoma de ovário com fenótipo de multirresistência) e NCI-H460 (carcinoma pulmonar). Conclusão: Esses resultados sugerem um promissor efeito auxiliar do zoledronato para o tratamento de alguns tipos de tumores; estudos complementares in vitro e in vivo são necessários para a validação dessa hipótese.


Introduction: Bisphosphonates are used in the treatment of bone diseases such as osteoporosis and bone metastases, because of their ability to inhibit bone resorption. There is evidence that high-potency bisphosphonates, such as zoledronate, are useful in the treatment of other malignancies because they have an antiproliferative effect. Objective:To evaluate the antiproliferative activity of zoledronate in different tumor cell lines. Method: This was an in vitro study in which nine human cell lines (U251, MCF7, NCI/ ADR-RES, 786-0, NCI-H460, PC-3, OVCAR-3, HT29, K-562, and HaCaT) were treated with of 0.12, 1.2, 12, and 120 µM of zoledronate, their proliferative activity being evaluated 48 h later with sulforhodamine B assay. Results: At the 12 µM and 120 µM doses, zoledronate effectively reduced the proliferation of 786-0 (renal carcinoma) cells by 50% and 100%, respectively. At the highest concentration (120 µM), zoledronate had a cytostatic effect (50% reduction in cell proliferation) on HaCaT (non-tumor human keratinocyte), HT-29 (colon carcinoma), NCI-ADR/ RES (multidrug-resistant ovarian adenocarcinoma), and NCI-H460 (lung carcinoma) cells. Conclusion: These results suggest a promising auxiliary effect of zoledronate for the treatment of some tumors. Further in vitro and in vivo studies are needed in order to test that hypothesis.


Introducción: Los bisfosfonatos son fármacos utilizados para el tratamiento de enfermedades óseas, como la osteoporosis y metástasis óseas debido a su mecanismo de acción, que consiste en la disminución del proceso de reabsorción del hueso. Otros estudios observaron que los bisfosfonatos de alta potencia, como el zoledronato, podrían ayudar en el tratamiento de otras enfermedades malignas debido a la promoción de un efecto antiproliferativo. Objetivo: Este estudio in vitro objetivó evaluar la actividad antiproliferativa de zoledronato en diferentes linajes de células tumorales. Método: Los nueve humano linajes (U251, MCF7, NCI / ADR-RES, 786-0, NCI-H460, PC-3, OVCAR-3, HT29, K-562 and HaCaT) se sometieron al tratamiento con las concentraciones de 0,12; 1,2; 12 y 120 µM de zoledronato y tuvieron su actividad proliferativa evaluada después de 48 horas utilizando el colorante sulforrodamina B. Resultados: Se comprobó que las concentraciones de 12 µM y 120 µM de zoledronato fueron efectivas para reducir en un 50% y un 100%, respectivamente, de la proliferación de las células 786-0 (carcinoma renal). La mayor concentración de zoledronato (120 µM) promovió un efecto citostático (reducción de la proliferación celular en un 50%) para las células HaCaT (queratinocito humano no tumoral), HT-29 (carcinoma de colon), NCI-ADR/RES (adenocarcinoma de ovário con fenótipo de multirresistencia) y NCI-H460 (carcinoma pulmonar). Conclusión: Estos resultados sugieren un prometedor efecto auxiliar del zoledronato para el tratamiento de algunos tumores; se requieren más estudios in vitro e in vivo para validar esta hipótesis


Assuntos
Humanos , Proliferação de Células/efeitos dos fármacos , Difosfonatos , Técnicas In Vitro , Células Tumorais Cultivadas/efeitos dos fármacos , Linhagem Celular Tumoral/efeitos dos fármacos
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