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1.
Life Sci ; 254: 117786, 2020 May 18.
Artigo em Inglês | MEDLINE | ID: mdl-32433918

RESUMO

AIMS: Ovarian cancer (OC) is the most lethal gynecological malignancies and many women develop chemoresistance associated with the inflammatory process. We investigated the effects of P-MAPA and IL-12 on the inflammatory and immune responses in a chemically-induced OC model. MAIN METHODS: OCs were induced with 7,12-dimethylbenz(a)anthracene into the ovarian bursa, and the animals were given P-MAPA (5 mg/kg bw., i.p., twice a week), or IL-12 (300 ng/kg bw., i.p., one a week) for 60 days, or both P-MAPA and IL-12. Immunohistochemistry, western blot, flow cytometry, and multiplex assay were used to examine the effectiveness of immunotherapies in OC. KEY FINDINGS: The combinatory therapy improved the general OC features, reducing inflammatory cells and adipocyte accumulation, in addition to revealing a soft and mobile tissue with no adherences and peritoneal implants. P-MAPA treatment increased the levels of TLR2, TLR4 and TRIF in OCs while decreasing the number of regulatory T (Treg) cells. Additionally, the association of P-MAPA with IL-12 significantly increased the number of CD4+ and CD8+ T effector cells in draining lymph nodes. Regarding the inflammatory mediators, P-MAPA enhanced the levels of the pro-inflammatory cytokine IL-17 while P-MAPA+IL-12 increased the levels of IL-1ß. Treatment with IL-12 enhanced the cytokine levels of IL-17, TNF-α, IL-1ß, and IL-2 in addition to the chemokine MIP-1α. SIGNIFICANCE: We conclude that P-MAPA upregulated TLR2 and TLR4 signaling, possibly activating the non-canonical pathway, while attenuating the tumor immunosuppression. Also, the combination of P-MAPA with IL-12 improves the antitumor immunoresponse, opening a new therapeutic approach for fighting OC.

2.
Eur J Pharm Biopharm ; 151: 162-170, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32311428

RESUMO

Bladder cancer is the fifth most common disease in the United States, and the treatment and alternatives for patients have not changed in the last decades. Silver nanoparticles (AgNP) have been used in the treatment of various cancer, mainly because of the antineoplastic activity; however, their use and the molecular mechanisms towards bladder cancer still unexplored. Therefore, this work aims to evaluate the in vitro and in vivo antitumoral mechanisms of biogenic silver nanoparticles synthesized from Fusarium sp. First, AgNP showed cytotoxicity in a dose- and time-response relationship and detailed analysis demonstrated the induction of cell death via apoptosis, also inhibiting cell migration and proliferation in bladder carcinoma cell line 5637. Next, it was evaluated the antitumoral activity of AgNP against non-muscle invasive bladder cancer (NMIBC). Bladder cancer was chemically induced with N-methyl-N-nitrosourea (MNU) on C57BL/6JUnib female mice and treated by intravesical route with AgNP concentrations of 0.5, 0.2, and 0.05 mg/mL. Finally, treatment with AgNP (0.05 mg/mL) led to 57.13% of tumor regression, with 14.28% of the animals showing normal urothelium, and 42.85% showing flat hyperplasia, considered to be a benign lesion. Overall, these findings demonstrated that AgNP might be a cost-effective alternative and promising candidate for the treatment of bladder cancer.

3.
Lipids Health Dis ; 19(1): 14, 2020 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-31996229

RESUMO

BACKGROUND: Altered lipid metabolism is an important characteristic of neoplastic cells, with androgens and growth factors being major regulatory agents of the lipid metabolism process. We investigated the effect of physical resistance training on lipid metabolism and apoptosis in the adult Wistar rat prostate. METHODS: Two experimental groups represented sedentary and physical resistance training. Three days per week for 13 weeks, rats performed jumps in water carrying a weight load strapped to their chests as part of a physical resistance exercise protocol. Two days after the last training session, rats were anesthetized and sacrificed for blood and prostate analysis. RESULTS: Physical exercise improved feeding efficiency, decreased weight gain, regulated the serum-lipid profile, and modulated insulin-like growth factor-1 (IGF-1) and free testosterone concentration. Furthermore, upregulation of cluster of differentiation 36 (CD36), sterol regulatory element binding protein-1 (SREBP-1), sterol regulatory element-binding protein cleavage-activating protein (SCAP), and reduced lysosome membrane protein (LIMPII) expression were also observed in the blood and prostates of trained rats. Consistent with these results, caspase-3 expression was upregulating and the BCL-2/Bax index ratio was decreased in trained rats relative to sedentary animals. CONCLUSIONS: In this work, physical resistance training can alter lipid metabolism and increase markers of apoptosis in the prostate, suggesting physical resistance training as a potential novel therapeutic strategy for treating prostate cancer.

4.
Life Sci ; 242: 117185, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31862453

RESUMO

Colorectal cancer (CRC) is a multifactorial syndrome that drives to uncontrollable cell division, genetic alterations, and functional alteration. In the present work, we evaluated the immunomodulatory properties of P-mapa, a compound extracted from Aspergillus oryzae fungus, versus Fluorouracil (5-FU) treatment in chemically induced CRC. CRC was induced by DMH in F344 rats. Animals of treated groups receive weekly 15 mg/Kg of 5-FU or 5 mg/Kg of P-mapa, over 10 weeks. Tissues were stained for aberrant crypt foci (ACF) counting and histopathology evaluation, immunostained for TLR4 pathways and quantified for TNFα Cytokine assay. DMH was efficient to induce hyperplastic lesions and ACF. Both treatments reduced significantly ACF formation and tumor aggressiveness. Immunohistochemistry for TLR4 signaling reveals that both treatments had no effect over the TLR4-NFκB signaling pathway. On the other hand, both succeed in increase interferon signaling, with activation of the TRIF-IRF3 pathway and consequently inducing IFNγ synthesis. The present results show the immunomodulatory properties of P-mapa in chemically induced CRC model. P-mapa induced a significant increase in Type-I IFNs synthesis and subsequently immune cell recruitment, resulting in an increase of IFNγ concentration in colorectal mucosa and its inhibitory effects over tumoral growth. In this scenario, P-mapa showed an interesting antitumoral effect by inhibiting tumor growth.


Assuntos
Antineoplásicos/uso terapêutico , Neoplasias do Colo/tratamento farmacológico , Fatores Imunológicos/uso terapêutico , Ácidos Linoleicos/uso terapêutico , Ácidos Oleicos/uso terapêutico , Transdução de Sinais/efeitos dos fármacos , Receptor 4 Toll-Like/metabolismo , Focos de Criptas Aberrantes/patologia , Animais , Biopolímeros/uso terapêutico , Neoplasias do Colo/metabolismo , Neoplasias do Colo/patologia , Ensaio de Imunoadsorção Enzimática , Fluoruracila/uso terapêutico , Masculino , Ratos , Ratos Endogâmicos F344 , Fator de Necrose Tumoral alfa/metabolismo
5.
Molecules ; 25(1)2019 Dec 18.
Artigo em Inglês | MEDLINE | ID: mdl-31861351

RESUMO

Immunotherapies have emerged as promising complementary treatments for ovarian cancer (OC), but its effective and direct role on OC cells is unclear. This study examined the combinatory effects of the protein aggregate magnesium-ammonium phospholinoleate-palmitoleate anhydride, known as P-MAPA, and the human recombinant interleukin-12 (hrIL-12) on cell migration/invasion, apoptosis, toll-like receptor (TLR)-mediated inflammation, and cytokine/chemokine profile in human OC cell line SKOV-3. P-MAPA and IL-12 showed cancer cell toxicity under low doses after 48 h. Although apoptosis/necrosis and the cell cycle were unchanged by the treatments, P-MAPA enhanced the sensitivity to paclitaxel (PTX) and P-MAPA associated with IL-12 significantly reduced the migratory potential and invasion capacity of SKOV-3 cells. P-MAPA therapy reduced TLR2 immunostaining and the myeloid differentiation factor 88 (MyD88), but not the TLR4 levels. Moreover, the combination of P-MAPA with IL-12 attenuated the levels of MyD88, interferon regulatory factor 3 (IRF3) and nuclear factor kappa B (NF-kB p65). The IL-12 levels were increased and P-MAPA stimulated the secretion of cytokines IL-3, IL-9, IL-10, and chemokines MDC/CCL22 and, regulated on activation, normal T cells expressed and secreted (RANTES)/CCL5. Conversely, combination therapy reduced the levels of IL-3, IL-9, IL-10, MDC/CCL22, and RANTES/CCL5. Collectively, P-MAPA and IL-12 reduce cell dynamics and effectively target the TLR-related downstream molecules, eliciting a protective effect against chemoresistance. P-MAPA also stimulates the secretion of anti-inflammatory molecules, possibly having an immune response in the OC microenvironment.


Assuntos
Mediadores da Inflamação/metabolismo , Interleucina-12/metabolismo , Ácidos Linoleicos/metabolismo , Ácidos Oleicos/metabolismo , Neoplasias Ovarianas/metabolismo , Receptores Toll-Like/metabolismo , Apoptose , Movimento Celular , Sobrevivência Celular/efeitos dos fármacos , Citocinas/metabolismo , Feminino , Humanos , Imunofenotipagem , Modelos Biológicos , Neoplasias Ovarianas/etiologia , Neoplasias Ovarianas/patologia , Neoplasias Ovarianas/terapia , Transdução de Sinais/efeitos dos fármacos
6.
Life Sci ; 237: 116895, 2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31610204

RESUMO

To evaluate the effect of a probiotic on the aggressiveness of a chemically induced colorectal tumor in rats. Twenty-five male Fisher 344 rats, 250 g, provided with feed and water ad libitum, were randomly divided into 5 groups (5 rats/group): GControl, no treatment; GTumor, tumor induction; GTumor+5FU, tumor induction, 5-Fluorouracil applied; GTumor+Prob, induction of the tumor, supplemented with probiotic; GTumor+5-FU+Prob, tumor induction, 5-Fluorouracil applied, supplemented with probiotic. For tumor induction 20 mg/kg of 1,2-dimethylhydrazine was applied intraperitoneally over 4 weeks, followed by an interval of 15 days, and then repeated for a further 4 weeks. Five weeks after the final dose of the carcinogen, treatment was initiated with 5-Fluorouracil (15 mg/kg, intraperitoneally/week) and a commercial probiotic (1 × 109 CFU, daily/gavage). Data were analyzed by One Way Variance Analysis and means compared by Dunnett's test. GraphPad Prism statistical software was used. The histopathological analyzes were evaluated by the chi-square test. A 5% type-I error was considered statistically significant. Compared with the GTumor, the GTumor+Prob (p < 0.0373) and GTumor+5-FU+Prob (p < 0.0003) demonstrated an attenuated effect on the aggressiveness of the colorectal tumor, with a reduction in the count of Aberrant Crypt foci; and a lower percentage of malignant neoplastic lesions in the GTumor+Prob (40% low grade tubular adenoma, 40% carcinoma in situ, 20% low grade adenocarcinoma) and GTumor+5-FU+Prob (40% low grade tubular adenoma and 60% carcinoma in situ). Probiotic supplementation has the potential to decrease the formation of aberrant crypts and ameliorate tumor malignancy, enhancing the antitumor effect of 5-Fluorouracil chemotherapy in colic segments.


Assuntos
Neoplasias Colorretais/prevenção & controle , Suplementos Nutricionais , Dimetilidrazinas/toxicidade , Probióticos/administração & dosagem , Animais , Carcinógenos/toxicidade , Neoplasias Colorretais/induzido quimicamente , Neoplasias Colorretais/patologia , Masculino , Ratos , Ratos Endogâmicos F344
7.
Horm Cancer ; 9(3): 175-187, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29363091

RESUMO

Use of drug combinations that target different pathways involved in the development and progression of prostate cancer (PCa) has emerged as an alternative to overcome the resistance caused by drug monotherapies. The antiandrogen abiraterone acetate and the PI3K/Akt inhibitor NVP-BEZ235 (BEZ235) may be suitable options for the prevention of drug resistance and the inhibition of PCa progression. The aim of the present study was to evaluate whether abiraterone acetate and BEZ235 achieve superior therapeutic effects to either drug administered as monotherapy, in the early stages of PCa in an androgen-dependent system. Our study showed that each drug might impair tumor growth by reducing proliferation and increasing cell death when administered as monotherapy. However, tumor growth continued to progress with each drug monotherapy and some important side effects were related to BEZ. Conversely, when used in combination, the drugs impaired the inflammatory response, decreased hyperplastic lesions, and blocked tumor progression from premalignant to a malignant stage. Our data showed that the strategy to block the androgenic and PI3K/AKT/mTOR pathway is an effective therapeutic option and should be investigated including distinct PI3K pathway inhibitors.


Assuntos
Acetato de Abiraterona/uso terapêutico , Antagonistas de Androgênios/uso terapêutico , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Imidazóis/uso terapêutico , Neoplasias da Próstata/tratamento farmacológico , Inibidores de Proteínas Quinases/uso terapêutico , Quinolinas/uso terapêutico , Androgênios/metabolismo , Animais , Carcinogênese , Modelos Animais de Doenças , Sinergismo Farmacológico , Humanos , Masculino , Ratos , Ratos Endogâmicos F344 , Transdução de Sinais , Células Tumorais Cultivadas
8.
J Ovarian Res ; 11(1): 8, 2018 Jan 17.
Artigo em Inglês | MEDLINE | ID: mdl-29343281

RESUMO

BACKGROUND: Toll-like receptors (TLRs) are transmembrane proteins expressed on the surface of ovarian cancer (OC) and immune cells. Identifying the specific roles of the TLR-mediated signaling pathways in OC cells is important to guide new treatments. Because immunotherapies have emerged as the adjuvant treatment for patients with OC, we investigated the effect of a promising immunotherapeutic strategy based on protein aggregate magnesium-ammonium phospholinoleate-palmitoleate anhydride (P-MAPA) combined with cisplatin (CIS) on the TLR2 and TLR4 signaling pathways via myeloid differentiation factor 88 (MyD88) and TLR-associated activator of interferon (TRIF) in an in vivo model of OC. METHODS: Tumors were chemically induced by a single injection of 100 µg of 7,12-dimethylbenz(a)anthracene (DMBA) directly under the left ovarian bursa in Fischer 344 rats. After the rats developed serous papillary OC, they were given P-MAPA, CIS or the combination P-MAPA+CIS as therapies. To understand the effects of the treatments, we assessed the tumor size, histopathology, and the TLR2- and TLR4-mediated inflammatory responses. RESULTS: Although CIS therapy was more effective than P-MAPA in reducing the tumor size, P-MAPA immunotherapy significantly increased the expressions of TLR2 and TLR4. More importantly, the combination of P-MAPA with CIS showed a greater survival rate compared to CIS alone, and exhibited a significant reduction in tumor volume compared to P-MAPA alone. The combination therapy also promoted the increase in the levels of the following OC-related proteins: TLR4, MyD88, TRIF, inhibitor of phosphorylated NF-kB alpha (p-IkBα), and nuclear factor kappa B (NF-kB p65) in both cytoplasmic and nuclear sites. While P-MAPA had no apparent effect on tumor necrosis factor alpha (TNF-α) and interleukin (IL)-6, it seems to increase interferon-γ (IFN-γ), which may induce the Thelper (Th1)-mediated immune response. CONCLUSION: Collectively, our results suggest that P-MAPA immunotherapy combined with cisplatin could be considered an important therapeutic strategy against OC cells based on signaling pathways activated by TLR4.


Assuntos
Cistadenocarcinoma Seroso/metabolismo , Cistadenocarcinoma Seroso/patologia , Neoplasias Ovarianas/metabolismo , Neoplasias Ovarianas/patologia , Transdução de Sinais/efeitos dos fármacos , Receptor 4 Toll-Like/metabolismo , Animais , Protocolos de Quimioterapia Combinada Antineoplásica/efeitos adversos , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Biomarcadores , Linhagem Celular Tumoral , Cisplatino/administração & dosagem , Cistadenocarcinoma Seroso/tratamento farmacológico , Cistadenocarcinoma Seroso/imunologia , Citocinas/metabolismo , Modelos Animais de Doenças , Feminino , Humanos , Imuno-Histoquímica , Ácidos Linoleicos/administração & dosagem , NF-kappa B , Gradação de Tumores , Compostos Organofosforados/administração & dosagem , Neoplasias Ovarianas/tratamento farmacológico , Neoplasias Ovarianas/imunologia , Ratos , Receptor 2 Toll-Like/metabolismo , Carga Tumoral , Ensaios Antitumorais Modelo de Xenoenxerto
9.
MAbs ; 10(1): 46-54, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-28581886

RESUMO

Despite fast advances in genomics and proteomics, monoclonal antibodies (mAbs) are still a valuable tool for areas such as the evolution of basic research in stem cells and cancer, for immunophenotyping cell populations, diagnosing and prognosis of diseases, and for immunotherapy. To summarize different subtractive immunization approaches successfully used for the production of highly specific antibodies, we identified scientific articles in NCBI PubMed using the following search terms: subtractive immunization, monoclonal antibody, tolerization, neonatal, high-zone tolerance, masking immunization. Patent records were also consulted. From the list of results, we included all available reports, from 1985 to present, that used any enhanced immunization technique to produce either polyclonal or monoclonal antibodies. Our examination yielded direct evidence that these enhanced immunization techniques are efficient in obtaining specific antibodies to rare epitopes, with different applications, such as to identify food contaminants or tumor cells.


Assuntos
Anticorpos Monoclonais/biossíntese , Anticorpos Monoclonais/imunologia , Especificidade de Anticorpos , Antígenos/administração & dosagem , Antígenos/imunologia , Imunização/métodos , Epitopos Imunodominantes/imunologia , Animais , Animais Recém-Nascidos , Humanos , Tolerância Imunológica , Esquemas de Imunização
10.
Int Urol Nephrol ; 49(4): 597-605, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28181115

RESUMO

OBJECTIVES: To evaluate the effect of implanted S-nitrosoglutathione (GSNO) coating polypropylene mesh in foreign-body response of rats. METHODS: Thirty female rats underwent to subcutaneous implant of five polypropylene (PP) fragments: uncoated PP (control); PP polyvinylalcohol (PVA) coated and PP PVA + GSNO (1, 10 and 70 mMol) coated. After euthanasia (4 and 30 days), eight slides were prepared from each animal: hematoxylin-eosin (inflammatory response); unstained (birefringence collagen evaluation); TUNEL technique (apoptosis); and five for immunohistochemical processing: CD-31 (angiogenesis), IL-1 and TNF-α (proinflammatory cytokynes), iNOS (NO synthesis) and MMP-2 (collagen metabolism). The inflammation area, birefringence index, apoptotic index, immunoreactivity and vessel density were objectively measured. RESULTS: Inflammatory reaction area at 4 days was 11.3, 15.2, 25.1, 17.1 and 19.3% of pure PP, PVA, GSNO 1, 10 and 70 mM, respectively, p = 0.0006 (PP × Others). At 30 days lower inflammatory area was observed in GSNO 10 and 70 mM compared to pure PP (5.3, 5.2 and 11.1%, respectively, p = 0.0001). Vessel density was higher for GSNO 1 mM (25.5%) compared to pure PP (19.6%) at 30 days only, p = 0.0081. Apoptotic index at 4 days was lower for GSNO 1 mM (49.3%) than pure PVA (60.6%), p = 0.0124. GSNO 10 and 70 mM reduced their apoptotic index at 30 days compared to 4 days (49.9 vs. 36.9 and 59.1 vs. 47.5%, respectively, p = 0.0397). Birefringence index, IL-1, TNF, MMP-2 and iNOS were not different. CONCLUSIONS: Depending on concentrations, GSNO can increase angiogenesis, reduce inflammation and apoptosis compared to pure PP, without impact on cytokine, collagen organization/metabolism and endogenous NO synthesis.


Assuntos
Reação a Corpo Estranho , Neovascularização Fisiológica , Doadores de Óxido Nítrico/farmacologia , Óxido Nítrico/biossíntese , S-Nitrosoglutationa/farmacologia , Telas Cirúrgicas , Animais , Apoptose/efeitos dos fármacos , Feminino , Reação a Corpo Estranho/etiologia , Reação a Corpo Estranho/metabolismo , Reação a Corpo Estranho/patologia , Interleucina-1/análise , Metaloproteinase 2 da Matriz/análise , Neovascularização Fisiológica/efeitos dos fármacos , Neovascularização Fisiológica/fisiologia , Óxido Nítrico Sintase Tipo II/análise , Molécula-1 de Adesão Celular Endotelial a Plaquetas/análise , Polipropilenos , Álcool de Polivinil/farmacologia , Ratos , Ratos Wistar , Telas Cirúrgicas/efeitos adversos , Fator de Necrose Tumoral alfa/análise
11.
BJU Int ; 119(6): 948-954, 2017 06.
Artigo em Inglês | MEDLINE | ID: mdl-28093890

RESUMO

OBJECTIVE: To study a novel penile reinnervation technique using four sural nerve grafts and end-to-side neurorraphies connecting bilaterally the femoral nerve and the cavernous corpus and the femoral nerve and the dorsal penile nerves. PATIENTS AND METHODS: Ten patients (mean [± sd; range] age 60.3 [± 4.8; 54-68] years), who had undergone radical prostatectomy (RP) at least 2 years previously, underwent penile reinnervation in the present study. Four patients had undergone radiotherapy after RP. All patients reported satisfactory sexual activity prior to RP. The surgery involved bridging of the femoral nerve to the dorsal nerve of the penis and the inner part of the corpus cavernosum with sural nerve grafts and end-to-side neurorraphies. Patients were evaluated using the International Index of Erectile Function (IIEF) questionnaire and pharmaco-penile Doppler ultrasonography (PPDU) preoperatively and at 6, 12 and 18 months postoperatively, and using a Clinical Evolution of Erectile Function (CEEF) questionnaire, administered after 36 months. RESULTS: The IIEF scores showed improvements with regard to erectile dysfunction (ED), satisfaction with intercourse and general satisfaction. Evaluation of PPDU velocities did not reveal any difference between the right and left sides or among the different time points. The introduction of nerve grafts neither caused fibrosis of the corpus cavernosum, nor reduced penile vascular flow. CEEF results showed that sexual intercourse began after a mean of 13.7 months with frequency of sexual intercourse varying from once daily to once monthly. Acute complications were minimal. The study was limited by the small number of cases. CONCLUSIONS: A total of 60% of patients were able to achieve full penetration, on average, 13 months after reinnervation surgery. Patients previously submitted to radiotherapy had slower return of erectile function. We conclude that penile reinnervation surgery is a viable technique, with effective results, and could offer a new treatment method for ED after RP.


Assuntos
Disfunção Erétil/etiologia , Disfunção Erétil/cirurgia , Nervo Femoral/cirurgia , Pênis/inervação , Pênis/cirurgia , Complicações Pós-Operatórias/etiologia , Complicações Pós-Operatórias/cirurgia , Prostatectomia/efeitos adversos , Nervo Sural/transplante , Idoso , Sistema Nervoso Autônomo/cirurgia , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Procedimentos Neurocirúrgicos/métodos , Prostatectomia/métodos , Fatores de Tempo , Procedimentos Cirúrgicos Urológicos Masculinos/métodos
12.
Int. braz. j. urol ; 42(5): 942-954, Sept.-Oct. 2016. tab, graf
Artigo em Inglês | LILACS-Express | ID: lil-796874

RESUMO

ABSTRACT The present study describes the histopathological and molecular effects of P-MAPA (Protein aggregate magnesium-ammonium phospholinoleate-palmitoleate anhydride) intravesical immunotherapy combined with systemic doxorubicin or cisplatin for treatment of non-muscle invasive bladder cancer (NMIBC) in an appropriate animal model. Our results showed an undifferentiated tumor, characterizing a tumor invading mucosa or submucosa of the bladder wall (pT1) and papillary carcinoma in situ (pTa) in the Cancer group. The histopathological changes were similar between the combined treatment with intravesical P-MAPA plus systemic Cisplatin and P-MAPA immunotherapy alone, showing decrease of urothelial neoplastic lesions progression and histopathological recovery in 80% of the animals. The animals treated systemically with cisplatin or doxorubicin singly, showed 100% of malignant lesions in the urinary bladder. Furthemore, the combined treatment with P-MAPA and Doxorubicin showed no decrease of urothelial neoplastic lesions progression and histopathological recovery. Furthermore, Akt, PI3K, NF-kB and VEGF protein levels were significantly lower in intravesical P-MAPA plus systemic cisplatin and in intravesical P-MAPA alone treatments than other groups. In contrast, PTEN protein levels were significantly higher in intravesical P-MAPA plus systemic cisplatin and in intravesical P-MAPA alone treatments. Thus, it could be concluded that combination of intravesical P-MAPA immunotherapy and systemic cisplatin in the NMIBC animal model was effective, well tolerated and showed no apparent signs of antagonism between the drugs. In addition, intravesical P-MAPA immunotherapy may be considered as a valuable option for treatment of BCG unresponsive patients that unmet the criteria for early cystectomy.

13.
BMC Cancer ; 16: 422, 2016 07 07.
Artigo em Inglês | MEDLINE | ID: mdl-27389279

RESUMO

BACKGROUND: The new modalities for treating patients with non-muscle invasive bladder cancer (NMIBC) for whom BCG (Bacillus Calmette-Guerin) has failed or is contraindicated are recently increasing due to the development of new drugs. Although agents like mitomycin C and BCG are routinely used, there is a need for more potent and/or less-toxic agents. In this scenario, a new perspective is represented by P-MAPA (Protein Aggregate Magnesium-Ammonium Phospholinoleate-Palmitoleate Anhydride), developed by Farmabrasilis (non-profit research network). This study detailed and characterized the mechanisms of action of P-MAPA based on activation of mediators of Toll-like Receptors (TLRs) 2 and 4 signaling pathways and p53 in regulating angiogenesis and apoptosis in an animal model of NMIBC, as well as, compared these mechanisms with BCG treatment. RESULTS: Our results demonstrated the activation of the immune system by BCG (MyD88-dependent pathway) resulted in increased inflammatory cytokines. However, P-MAPA intravesical immunotherapy led to distinct activation of TLRs 2 and 4-mediated innate immune system, resulting in increased interferons signaling pathway (TRIF-dependent pathway), which was more effective in the NMIBC treatment. Interferon signaling pathway activation induced by P-MAPA led to increase of iNOS protein levels, resulting in apoptosis and histopathological recovery. Additionally, P-MAPA immunotherapy increased wild-type p53 protein levels. The increased wild-type p53 protein levels were fundamental to NO-induced apoptosis and the up-regulation of BAX. Furthermore, interferon signaling pathway induction and increased p53 protein levels by P-MAPA led to important antitumor effects, not only suppressing abnormal cell proliferation, but also by preventing continuous expansion of tumor mass through suppression of angiogenesis, which was characterized by decreased VEGF and increased endostatin protein levels. CONCLUSIONS: Thus, P-MAPA immunotherapy could be considered an important therapeutic strategy for NMIBC, as well as, opens a new perspective for treatment of patients that are refractory or resistant to BCG intravesical therapy.


Assuntos
Fatores Imunológicos/administração & dosagem , Ácidos Linoleicos/administração & dosagem , Neovascularização Patológica/tratamento farmacológico , Compostos Organofosforados/administração & dosagem , Receptores Toll-Like/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Neoplasias da Bexiga Urinária/tratamento farmacológico , Administração Intravesical , Animais , Vacina BCG/administração & dosagem , Vacina BCG/farmacologia , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Fatores Imunológicos/farmacologia , Imunoterapia/métodos , Ácidos Linoleicos/farmacologia , Invasividade Neoplásica , Neoplasias Experimentais , Neovascularização Patológica/metabolismo , Compostos Organofosforados/farmacologia , Ratos , Regulação para Cima , Neoplasias da Bexiga Urinária/irrigação sanguínea , Neoplasias da Bexiga Urinária/metabolismo
14.
Int Braz J Urol ; 42(3): 585-93, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27286125

RESUMO

OBJECTIVES: To describe acute and sub acute aspects of histological and immunohistochemical response to PP implant in a rat subcutaneous model based on objective methods. MATERIALS AND METHODS: Thirty rats had a PP mesh subcutaneously implanted and the same dissection on the other side of abdomen but without mesh (sham). The animals were euthanized after 4 and 30 days. Six slides were prepared using the tissue removed: one stained with hematoxylin-eosin (inflammation assessment); one unstained (birefringence evaluation) and four slides for immunohistochemical processing: IL-1 and TNF-α (pro-inflammatory cytokines), MMP-2 (collagen metabolism) and CD-31 (angiogenesis). The area of inflammation, the birefringence index, the area of immunoreactivity and the number of vessels were objectively measured. RESULTS: A larger area of inflammatory reaction was observed in PP compared to sham on the 4th and on the 30th day (p=0.0002). After 4 days, PP presented higher TNF (p=0.0001) immunoreactivity than sham and no differences were observed in MMP-2 (p=0.06) and IL-1 (p=0.08). After 30 days, a reduction of IL-1 (p=0.010) and TNF (p=0.016) for PP and of IL-1 (p=0.010) for sham were observed. Moreover, area of MMP-2 immunoreactivity decreased over time for PP group (p=0.018). Birefringence index and vessel counting showed no differences between PP and sham (p=0.27 and p=0.58, respectively). CONCLUSIONS: The implantation of monofilament and macroporous polypropylene in the subcutaneous of rats resulted in increased inflammatory activity and higher TNF production in the early post implant phase. After 30 days, PP has similar cytokines immunoreactivity, vessel density and extracellular matrix organization.


Assuntos
Reação a Corpo Estranho/induzido quimicamente , Reação a Corpo Estranho/etiologia , Reação a Corpo Estranho/patologia , Polipropilenos/efeitos adversos , Tela Subcutânea/patologia , Telas Cirúrgicas/efeitos adversos , Animais , Materiais Biocompatíveis/efeitos adversos , Birrefringência , Celulite (Flegmão)/etiologia , Celulite (Flegmão)/patologia , Colágeno/análise , Colágeno/metabolismo , Feminino , Imuno-Histoquímica , Interleucina-1/análise , Interleucina-1/metabolismo , Teste de Materiais , Metaloproteinase 2 da Matriz/análise , Metaloproteinase 2 da Matriz/metabolismo , Molécula-1 de Adesão Celular Endotelial a Plaquetas/análise , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Ratos Wistar , Reprodutibilidade dos Testes , Fatores de Tempo , Fator de Necrose Tumoral alfa/análise , Fator de Necrose Tumoral alfa/metabolismo
15.
Int. braz. j. urol ; 42(3): 585-593, tab, graf
Artigo em Inglês | LILACS-Express | ID: lil-785738

RESUMO

ABSTRACT Objectives To describe acute and sub acute aspects of histological and immunohistochemical response to PP implant in a rat subcutaneous model based on objective methods. Materials and Methods Thirty rats had a PP mesh subcutaneously implanted and the same dissection on the other side of abdomen but without mesh (sham). The animals were euthanized after 4 and 30 days. Six slides were prepared using the tissue removed: one stained with hematoxylin-eosin (inflammation assessment); one unstained (birefringence evaluation) and four slides for immunohistochemical processing: IL-1 and TNF-α (pro-inflammatory cytokines), MMP-2 (collagen metabolism) and CD-31 (angiogenesis). The area of inflammation, the birefringence index, the area of immunoreactivity and the number of vessels were objectively measured. Results A larger area of inflammatory reaction was observed in PP compared to sham on the 4th and on the 30th day (p=0.0002). After 4 days, PP presented higher TNF (p=0.0001) immunoreactivity than sham and no differences were observed in MMP-2 (p=0.06) and IL-1 (p=0.08). After 30 days, a reduction of IL-1 (p=0.010) and TNF (p=0.016) for PP and of IL-1 (p=0.010) for sham were observed. Moreover, area of MMP-2 immunoreactivity decreased over time for PP group (p=0.018). Birefringence index and vessel counting showed no differences between PP and sham (p=0.27 and p=0.58, respectively). Conclusions The implantation of monofilament and macroporous polypropylene in the subcutaneous of rats resulted in increased inflammatory activity and higher TNF production in the early post implant phase. After 30 days, PP has similar cytokines immunoreactivity, vessel density and extracellular matrix organization.

16.
Int Braz J Urol ; 42(5): 942-954, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-24893914

RESUMO

The present study describes the histopathological and molecular effects of P-MAPA (Protein aggregate magnesium-ammonium phospholinoleate-palmitoleate anhydride) intravesical immunotherapy combined with systemic doxorubicin or cisplatin for treatment of non-muscle invasive bladder cancer (NMIBC) in an appropriate animal model. Our results showed an undifferentiated tumor, characterizing a tumor invading mucosa or submucosa of the bladder wall (pT1) and papillary carcinoma in situ (pTa) in the Cancer group. The histopathological changes were similar between the combined treatment with intravesical P-MAPA plus systemic Cisplatin and P-MAPA immunotherapy alone, showing decrease of urothelial neoplastic lesions progression and histopathological recovery in 80% of the animals. The animals treated systemically with cisplatin or doxorubicin singly, showed 100% of malignant lesions in the urinary bladder. Furthemore, the combined treatment with P-MAPA and Doxorubicin showed no decrease of urothelial neoplastic lesions progression and histopathological recovery. Furthermore, Akt, PI3K, NF-kB and VEGF protein levels were significantly lower in intravesical P-MAPA plus systemic cisplatin and in intravesical P-MAPA alone treatments than other groups. In contrast, PTEN protein levels were significantly higher in intravesical P-MAPA plus systemic cisplatin and in intravesical P-MAPA alone treatments. Thus, it could be concluded that combination of intravesical P-MAPA immunotherapy and systemic cisplatin in the NMIBC animal model was effective, well tolerated and showed no apparent signs of antagonism between the drugs. In addition, intravesical P-MAPA immunotherapy may be considered as a valuable option for treatment of BCG unresponsive patients that unmet the criteria for early cystectomy.


Assuntos
Antineoplásicos/uso terapêutico , Carcinoma/terapia , Cisplatino/uso terapêutico , Doxorrubicina/uso terapêutico , Imunoterapia/métodos , Proteínas de Membrana/uso terapêutico , Neoplasias da Bexiga Urinária/terapia , Administração Intravesical , Animais , Vacina BCG , Western Blotting , Carcinoma/patologia , Terapia Combinada , Progressão da Doença , Feminino , Modelos Animais , NF-kappa B/análise , PTEN Fosfo-Hidrolase/análise , Fosfatidilinositol 3-Quinases/análise , Proteínas Proto-Oncogênicas c-akt/análise , Ratos Endogâmicos F344 , Reprodutibilidade dos Testes , Resultado do Tratamento , Neoplasias da Bexiga Urinária/patologia , Fator A de Crescimento do Endotélio Vascular/análise
17.
Int Braz J Urol ; 41(5): 849-58, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26689510

RESUMO

INTRODUCTION AND OBJECTIVES: Reactive Stroma (RStr) is observed in many human cancers and is related to carcinogenesis. The objectives of the present study were to stablish a relationship of the RStr microenvironment with prostate cancer (Pca) through a morphological and molecular characterization, and to identify a possible relationship between RStr with worse prognosis factors and occurrence of malignant prostatic stem cells. MATERIALS AND METHODS: Forty prostatic samples were selected from men with Pca diagnosis submitted to radical prostatectomy; they were divided in two groups: Group-1 (n=20): samples without reactive stroma; Group-2 (n=20): samples of PCa with intense stroma reaction. Prostatic samples were evaluated for RStr intensity by Masson Trichromic stain and posteriorly submitted to histopathological and immunohistochemistry analysis for antigens: a-actin, vimentin, IGF-1, MMP-2, FGF-2, C-Myc, PSCA, AR, Era and ERß. RESULTS: Reactive stroma with intense desmoplastic reactivity was significantly more frequent in intermediate (Gleason 7, 3+4) and high grade tumors (Gleason 7, 4+3). The group with intense stromal reactivity showed significant higher levels of Vimentin, IGF-1, MMP-2, FGF-2, C-Myc, PSCA and ERa. CONCLUSIONS: It can be concluded that RStr may be a predictive marker of Pca progression, since it was associated with increase of growth factors, imbalance of androgen and estrogen receptors and presence of malign prostatic stem cells.


Assuntos
Adenocarcinoma/patologia , Células Epiteliais/patologia , Células-Tronco Neoplásicas/patologia , Neoplasias da Próstata/patologia , Células Estromais/patologia , Actinas/análise , Adenocarcinoma/química , Idoso , Idoso de 80 Anos ou mais , Antígenos de Neoplasias/análise , Biomarcadores Tumorais/análise , Proteínas de Ligação a DNA/análise , Progressão da Doença , Células Epiteliais/química , Receptor alfa de Estrogênio/análise , Fator 2 de Crescimento de Fibroblastos/análise , Proteínas Ligadas por GPI/análise , Humanos , Imuno-Histoquímica , Fator de Crescimento Insulin-Like I/análise , Masculino , Metaloproteinase 2 da Matriz/análise , Pessoa de Meia-Idade , Gradação de Tumores , Proteínas de Neoplasias/análise , Células-Tronco Neoplásicas/química , Neoplasias da Próstata/química , Células Estromais/química , Fatores de Transcrição/análise , Microambiente Tumoral , Vimentina/análise
18.
Int. braz. j. urol ; 41(5): 849-858, Sept.-Oct. 2015. tab, graf
Artigo em Inglês | LILACS | ID: lil-767051

RESUMO

ABSTRACT Introduction and Objectives: Reactive Stroma (RStr) is observed in many human cancers and is related to carcinogenesis. The objectives of the present study were to stablish a relationship of the RStr microenvironment with prostate cancer (Pca) through a morphological and molecular characterization, and to identify a possible relationship between RStr with worse prognosis factors and occurrence of malignant prostatic stem cells. Materials and Methods: Forty prostatic samples were selected from men with Pca diagnosis submitted to radical prostatectomy; they were divided in two groups: Group-1 (n=20): samples without reactive stroma; Group-2 (n=20): samples of PCa with intense stroma reaction. Prostatic samples were evaluated for RStr intensity by Masson Trichromic stain and posteriorly submitted to histopathological and immunohistochemistry analysis for antigens: α-actin, vimentin, IGF-1, MMP-2, FGF-2, C-Myc, PSCA, AR, Erα and ERβ. Results: Reactive stroma with intense desmoplastic reactivity was significantly more frequent in intermediate (Gleason 7, 3+4) and high grade tumors (Gleason 7, 4+3). The group with intense stromal reactivity showed significant higher levels of Vimentin, IGF-1, MMP-2, FGF-2, C-Myc, PSCA and ERα. Conclusions: It can be concluded that RStr may be a predictive marker of Pca progression, since it was associated with increase of growth factors, imbalance of androgen and estrogen receptors and presence of malign prostatic stem cells.


Assuntos
Idoso , Idoso de 80 Anos ou mais , Humanos , Masculino , Pessoa de Meia-Idade , Adenocarcinoma/patologia , Células Epiteliais/patologia , Células-Tronco Neoplásicas/patologia , Neoplasias da Próstata/patologia , Células Estromais/patologia , Actinas/análise , Adenocarcinoma/química , Antígenos de Neoplasias/análise , Biomarcadores Tumorais/análise , Progressão da Doença , Proteínas de Ligação a DNA/análise , Células Epiteliais/química , Receptor alfa de Estrogênio/análise , /análise , Proteínas Ligadas por GPI/análise , Imuno-Histoquímica , Fator de Crescimento Insulin-Like I/análise , /análise , Gradação de Tumores , Proteínas de Neoplasias/análise , Células-Tronco Neoplásicas/química , Neoplasias da Próstata/química , Células Estromais/química , Microambiente Tumoral , Fatores de Transcrição/análise , Vimentina/análise
19.
Int J Clin Exp Pathol ; 8(5): 4427-43, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26191134

RESUMO

The present study describes the role of the ubiquitin ligase Siah-2 and corepressor N-CoR in controlling androgen receptor (AR) and estrogen receptors (ERα and ERß) signaling in an appropriate animal model (Fischer 344 female rats) of non-muscle invasive bladder cancer (NMIBC), especially under conditions of anti-androgen therapy with flutamide. Furthermore, this study describes the mechanisms of a promising therapeutic alternative for NMIBC based on Protein aggregate magnesium-ammonium phospholinoleate-palmitoleate anhydride (P-MAPA) intravesical immunotherapy combined with flutamide, involving the interaction among steroid hormone receptors, their regulators and Toll-like receptors (TLRs). Our results demonstrated that increased Siah-2 and AR protein levels and decreased N-CoR, cytochrome P450 (CYP450) and estrogen receptors levels played a critical role in the urothelial carcinogenesis, probably leading to escape of urothelial cancer cells from immune system attack. P-MAPA immunotherapy led to distinct activation of innate immune system TLRs 2 and 4-mediated, resulting in increase of interferon signaling pathway, which was more effective in recovering the immunosuppressive tumor immune microenvironment and in recovering the bladder histology features than BCG (Bacillus Calmette-Guerin) treatments. The AR blockade therapy was important in the modulating of downstream molecules of TLR2 and TLR4 signaling pathway, decreasing the inflammatory cytokines signaling and enhancing the interferon signaling pathway when associated with P-MAPA. Taken together, the data obtained suggest that interferon signaling pathway activation and targeting AR and Siah-2 signals by P-MAPA intravesical immunotherapy alone and/ or in combination with AR blockade may provide novel therapeutic approaches for NMIBC.


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Imunoterapia/métodos , Neoplasias da Bexiga Urinária/patologia , Administração Intravesical , Antagonistas de Androgênios/administração & dosagem , Animais , Western Blotting , Modelos Animais de Doenças , Feminino , Flutamida/administração & dosagem , Ácidos Linoleicos/administração & dosagem , Coativadores de Receptor Nuclear/metabolismo , Compostos Organofosforados/administração & dosagem , Ratos , Ratos Endogâmicos F344 , Receptores Androgênicos/metabolismo , Ubiquitina-Proteína Ligases/metabolismo
20.
World J Urol ; 33(3): 413-9, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24871424

RESUMO

OBJECTIVE: The objective of the study was to illustrate the applicability and significance of the novel Lewis urothelial cancer model compared to the classic Fisher 344. METHODS: Fischer 344 and Lewis females rats, 7 weeks old, were intravesical instilled N-methyl-N-nitrosourea 1.5 mg/kg every other week for a total of four doses. After 15 weeks, animals were sacrificed and bladders analyzed: histopathology (tumor grade and stage), immunohistochemistry (apoptotic and proliferative indices) and blotting (Toll-like receptor 2-TLR2, Uroplakin III-UP III and C-Myc). Control groups received placebo. RESULTS: There were macroscopic neoplastic lesions in 20 % of Lewis strain and 70 % of Fischer 344 strain. Lewis showed hyperplasia in 50 % of animals, normal bladders in 50 %. All Fischer 344 had lesions, 20 % papillary hyperplasia, 30 % dysplasia, 40 % neoplasia and 10 % squamous metaplasia. Proliferative and apoptotic indices were significantly lower in the Lewis strain (p < 0.01). The TLR2 and UP III protein levels were significantly higher in Lewis compared to Fischer 344 strain (70.8 and 46.5 % vs. 49.5 and 16.9 %, respectively). In contrast, C-Myc protein levels were significantly higher in Fischer 344 (22.5 %) compared to Lewis strain (13.7 %). CONCLUSIONS: The innovative Lewis carcinogen resistance urothelial model represents a new strategy for translational research. Preservation of TLR2 and UP III defense mechanisms might drive diverse urothelial phenotypes during carcinogenesis in differently susceptible individuals.


Assuntos
Carcinoma de Células de Transição/fisiopatologia , Modelos Animais de Doenças , Resistência à Doença/fisiologia , Receptor 2 Toll-Like/fisiologia , Neoplasias da Bexiga Urinária/fisiopatologia , Uroplaquina III/fisiologia , Animais , Apoptose/fisiologia , Carcinoma de Células de Transição/induzido quimicamente , Carcinoma de Células de Transição/patologia , Proliferação de Células/fisiologia , Feminino , Metilnitrosoureia/efeitos adversos , Fenótipo , Proteínas Proto-Oncogênicas c-myc/fisiologia , Ratos , Ratos Endogâmicos F344 , Ratos Endogâmicos Lew , Bexiga Urinária/patologia , Neoplasias da Bexiga Urinária/induzido quimicamente , Neoplasias da Bexiga Urinária/patologia
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