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1.
Pharm Dev Technol ; 25(1): 107-115, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31603017

RESUMO

In this study, mixed micelles of Soluplus® and TPGS were developed for co-administering docetaxel (DTX) and piperine (PIP) for exerting the synergistic effect, which increased the cytotoxicity and improved the anti-cancer activity in HepG2 cell lines compared to free DTX. These in vitro (MTT assay, intracellular uptake of micelles) and in vivo (pharmacokinetic study, immunostaining, TUNEL analysis) studies exhibited the advantages of co-delivery of anticancer drugs with Soluplus®/TPGS by mixed micelles and furthermore established that co-delivery of DTX and PIP via the mixed micelles of Soluplus®/TPGS could be a promising strategy for the treatment of liver cancer.

2.
Pharm Dev Technol ; 24(9): 1125-1132, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31305197

RESUMO

Clinically, co-delivery of chemotherapeutics has been limited by poor water-solubility and severe systemic toxicity. This study was aimed at integrating the merits of combination chemotherapy and mixed micellar technology and demonstrating the anticancer potential of doxorubicin (DOX) and dihydroartemisinin (DHA) co-loaded Soluplus®-TPGS mixed micellar system. In this study, physiochemically stable multidrug loaded mixed micelles were successfully prepared, encapsulation efficiencies of DOX and DHA were as high as 90%, and the average diameter of the micelles was 64.27 nm. The cellular uptake of DOX from the mixed micelles increased by 1.3 and 1.2 times for MCF-7 and MCF-7/ADR cell lines, respectively. The micelles were more cytotoxic than free DHA-DOX. Surprisingly, the co-loaded mixed micelles exhibited higher antitumor activity, while the systemic toxicity was reduced during the treatment. Therefore, the DOX and DHA mixed micelle might be a potential, effective, and less toxic drug-delivery system for cancer therapy.


Assuntos
Antibióticos Antineoplásicos/administração & dosagem , Antimaláricos/administração & dosagem , Artemisininas/administração & dosagem , Doxorrubicina/administração & dosagem , Polietilenoglicóis/química , Polivinil/química , Vitamina E/química , Antibióticos Antineoplásicos/farmacologia , Antimaláricos/farmacologia , Artemisininas/farmacologia , Neoplasias da Mama/tratamento farmacológico , Doxorrubicina/farmacologia , Portadores de Fármacos/química , Combinação de Medicamentos , Feminino , Humanos , Células MCF-7 , Micelas
3.
J Nanosci Nanotechnol ; 19(1): 188-193, 2019 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-30327021

RESUMO

AZ91 alloy with 1.0 wt.% (La, Ce) mischmetal (MM) addition was solution treated at 593 K for 4 h, 653 K for 2 h and 693 K for 8 h (T4), which was subsequently aged at 493 K for 5 h, 8 h and 11 h, respectively. Solid solution and aging treatment mainly led to the change of number of ß-Mg17Al12 phases and the size of mischmetal (MM) intermetallics. The relative potential difference is an index for local corrosion of alloys because the corrosion is driven by micro-galvanic coupling of the microstructure. In the present work, the relative Volta potential difference between the dominated intermetallic phase and Mg-matrix phase in the AZ91 alloy with (La, Ce) MM addition after heat treatment was measured by scanning Kelvin probe force microscopy (SKPFM). Then correlative changes of relative Volta potentials of intermetallic phases with solid-solution-treatment and subsequently aging treatment (T6) were inferred. The relative Volta potential difference of mischmetal intermetallic phases increased with the solid solution treatment. With the increase of aging time, the relative Volta potential difference of mischmetal intermetallic phases decreased. The relative Volta potential difference of intermetallic phases containing rare earth was ranked as: T6-11 h

4.
J Tissue Eng Regen Med ; 12(9): 1959-1971, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-30055109

RESUMO

Multidrug resistance (MDR) is one of the major obstacles to improving outcomes of chemotherapy in tumour patients. However, progress has been slow to overcome this phenomenon due to the limitations of current cell/tissue models in recapitulating MDR behaviour of tumour cells in vitro. To address this issue, a more pathologically relevant, three-dimensional (3D) culture of human breast cancer cells was developed by seeding the adriamycin-resistant cells MCF-7R in silk-collagen scaffolds. The cultures of the parental cell line MCF-7 served as controls. Distinct growth profiles of MCF-7R and MCF-7 cells were observed when they were cultured in the scaffolds in comparison with those in the monolayer culture, including cell proliferation, cellular aggregate formation, and expression of drug resistance-related genes/proteins. Moreover, the 3D cultures of these cell lines especially the cultures of MCF-7R exhibited a significantly enhanced drug resistance evidenced by their increased IC50 values to the anticancer drugs and improved drug efflux capability. An altered cell cycle distribution and improved percentage of breast cancer stem cell (BCSC)-like cells was also found in the present study. This might play an important role in promoting the drug-resistance production in those 3D cultures. Thus, we established improved 3D cultures of MDR human breast cancer. It would provide a robust tissue model for use to evaluate the efficacy of anticancer drugs, explore mechanisms of MDR, and enrich BCSCs in vitro.


Assuntos
Neoplasias da Mama/patologia , Resistência a Múltiplos Medicamentos , Resistencia a Medicamentos Antineoplásicos , Técnicas de Cultura de Tecidos/métodos , Neoplasias da Mama/genética , Neoplasias da Mama/ultraestrutura , Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Forma Celular/efeitos dos fármacos , Doxorrubicina/farmacologia , Resistência a Múltiplos Medicamentos/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Células MCF-7 , Células-Tronco Neoplásicas/efeitos dos fármacos , Células-Tronco Neoplásicas/metabolismo , Seda/química , Tecidos Suporte/química
5.
Drug Dev Ind Pharm ; 44(9): 1409-1416, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29718714

RESUMO

BACKGROUND: Although piperine can inhibit cells of tumors, the poor water solubility restricted its clinical application. This paper aimed to develop mixed micelles based on Soluplus® and D-α-tocopherol polyethylene glycol succinate (TPGS) to improve the aqueous solubility and anti-cancer effect. METHODS: Piperine-loaded mixed micelles were prepared using a thin-film hydration method, and their physicochemical properties were characterized. The cellular uptake of the micelles was confirmed by confocal laser scanning microscopy in A549 lung cancer cells and HepG2 liver cancer cells. In addition, cytotoxicity of the piperine mixed micelles was studied in A549 lung cancer cells and HepG2 liver cancer cells. Free piperine or piperine-loaded Soluplus®/TPGS mixed micelles were administered at an equivalent dose of piperine at 3.2 mg/kg via a single intravenous injection in the tail vain for the pharmacokinetic study in vivo. RESULTS: The diameter of piperine-loaded Soluplus®/TPGS (4:1) mixed micelles was about 61.9 nm and the zeta potential -1.16 ± 1.06 mV with 90.9% of drug encapsulation efficiency and 4.67% of drug-loading efficiency. Differential scanning calorimetry (DSC) studies confirmed that piperine is encapsulated by the Soluplus®/TPGS. The release results in vitro showed that the piperine-loaded Soluplus®/TPGS mixed micelles presented sustained release behavior compared to the free piperine. The mixed micelles exhibited better antitumor efficacy compared to free piperine and physical mixture against in A549 and HepG2 cells by MTT assay. The pharmacokinetic study revealed that the AUC of piperine-loaded mixed micelles was 2.56 times higher than that of piperine and the MRT for piperine-loaded mixed micelles was 1.2-fold higher than piperine (p < .05). CONCLUSION: The results of the study suggested that the piperine-loaded mixed micelles developed might be a potential nano-drug delivery system for cancer chemotherapy. These results demonstrated that piperine-loaded Soluplus®/TPGS mixed micelles are an effective strategy to deliver piperine for cancer therapy.


Assuntos
Alcaloides/química , Alcaloides/farmacologia , Benzodioxóis/química , Benzodioxóis/farmacologia , Neoplasias/tratamento farmacológico , Piperidinas/química , Piperidinas/farmacologia , Polietilenoglicóis/química , Alcamidas Poli-Insaturadas/química , Alcamidas Poli-Insaturadas/farmacologia , Polivinil/química , Vitamina E/química , Células A549 , Animais , Antineoplásicos/química , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Preparações de Ação Retardada/química , Preparações de Ação Retardada/farmacologia , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos/métodos , Células Hep G2 , Humanos , Masculino , Micelas , Ratos Sprague-Dawley , Solubilidade/efeitos dos fármacos
6.
Drug Dev Ind Pharm ; 43(7): 1197-1204, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28300426

RESUMO

BACKGROUND: Dioscin has shown cytotoxicity against cancer cells, but its poor solubility and stability have limited its clinical application. In this study, we designed mixed micelles composed of TPGS and Soluplus® copolymers entrapping the poorly soluble anticancer drug dioscin. METHOD: In order to improve the aqueous solubility and bioactivity of dioscin, TPGS/Soluplus® mixed micelles with an optimal ratio were prepared using a thin-film hydration method, and their physicochemical properties were characterized. Cellular cytotoxicity and uptake of the dioscin-loaded TPGS/Soluplus® mixed micelles were studied in MCF-7 breast cancer cells and A2780s ovarian cancer cells. The pharmacokinetics of free dioscin and dioscin-loaded TPGS/Soluplus® mixed micelles was studied in vivo in male Sprague-Dawley rats via a single intravenous injection in the tail vein. RESULTS: The average size of the optimized mixed micelle was 67.15 nm, with 92.59% drug encapsulation efficiency and 4.63% drug loading efficiency. The in vitro release profile showed that the mixed micelles presented sustained release behavior compared to the anhydrous ethanol solution of dioscin. In vitro cytotoxicity assays were conducted on human cancer cell lines including A2780s ovarian cancer cells and MCF-7 breast cancer cells. The mixed micelles exhibited better antitumor activity compared to free dioscin against all cell lines, which may benefit from the significant increase in the cellular uptake of dioscin from mixed micelles compared to free dioscin. The pharmacokinetic study showed that the mixed micelle formulation achieved a 1.3 times longer mean residual time (MRT) in circulation and a 2.16 times larger area under the plasma concentration-time curve (AUC) than the free dioscin solution. CONCLUSION: Our results suggest that the dioscin-loaded mixed micelles developed in this study might be a potential nano drug-delivery system for cancer chemotherapy.


Assuntos
Antineoplásicos/administração & dosagem , Antineoplásicos/farmacologia , Neoplasias da Mama/tratamento farmacológico , Diosgenina/análogos & derivados , Neoplasias Ovarianas/tratamento farmacológico , Poloxâmero/química , Polietilenoglicóis/administração & dosagem , Polivinil/administração & dosagem , Vitamina E/administração & dosagem , Animais , Antineoplásicos/química , Linhagem Celular Tumoral , Diosgenina/administração & dosagem , Diosgenina/química , Diosgenina/farmacologia , Feminino , Humanos , Células MCF-7 , Micelas , Polietilenoglicóis/química , Polivinil/química , Ratos Sprague-Dawley , Vitamina E/química
7.
J Colloid Interface Sci ; 485: 91-98, 2017 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-27657837

RESUMO

BACKGROUND: Nanosuspension is one of the most promising strategies to improve the oral bioavailability of insoluble drugs. The existing techniques applied to produce nanosuspensions are classified as "bottom-up" or "top-down" methods, or a combination of both. Curcumin (CUR), a Biopharmaceutics Classification System (BCS) class IV substance, is a promising drug candidate in view of its good bioactivity, but its use is limited due to its poor solubility and permeability. In the present study, CUR nanosuspensions were developed to enhance CUR oral bioavailability using a cost-effective method different from conventional techniques. RESULTS: The physicochemical properties of CUR nanosuspensions were characterized by dynamic light scattering (DLS) and transmission electron microscopy (TEM). The crystalline state of CUR in different nanosuspensions analyzed using differential scanning calorimeter (DSC) and X-ray diffraction analysis (PXRD) confirmed its amorphous state. In vitro dissolution degree of the prepared CUR nanosuspensions using TPGS or Brij78 as stabilizer was greatly increased. Pharmacokinetic studies demonstrated that the oral bioavailability of CUR was increased 3.18 and 3.7 times after administration of CUR/TPGS nanosuspensions or CUR/Brij78 nanosuspensions, when compared with the administration of CUR suspension. CONCLUSIONS: CUR nanosuspensions produced by our cost-effective method could improve its oral bioavailability. In addition, the low-cost and time-saving method reported here is highly suitable for a fast and inexpensive preparation.


Assuntos
Curcumina/farmacocinética , Nanoestruturas/química , Polietilenoglicóis/química , Vitamina E/química , Administração Oral , Animais , Disponibilidade Biológica , Dióxido de Carbono , Curcumina/química , Curcumina/economia , Estabilidade de Medicamentos , Masculino , Nanoestruturas/economia , Nanoestruturas/ultraestrutura , Ratos , Ratos Sprague-Dawley , Suspensões
8.
Oncotarget ; 7(33): 53254-53268, 2016 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-27449101

RESUMO

In this study, we examined the role of the miRNA miR-770-5p in cisplatin chemotherapy resistance in ovarian cancer (OVC) patients. miR-770-5p expression was reduced in platinum-resistant patients. Using a 6.128-fold in expression as the cutoff value, miR-770-5p expression served as a prognostic biomarker and predicted the response to cisplatin treatment and survival among OVC patients. Overexpression of miR-770-5p in vitro reduced survival in chemoresistant cell lines after cisplatin treatment. ERCC2, a target gene of miR-770-5p that participates in the NER system, was negatively regulated by miR-770-5p. siRNA-mediated silencing of ERCC2 reversed the inhibition of apoptosis resulting from miR-770-5p downreglation in A2780S cells. A comet assay confirmed that this restoration of cisplatin chemosensitivity was due to the inhibition of DNA repair. These findings suggest that endogenous miR-770-5p may function as an anti-oncogene and promote chemosensitivity in OVC, at least in part by downregulating ERCC2. miR-770-5p may therefore be a useful biomarker for predicting chemosensitivity to cisplatin in OVC patients and improve the selection of effective, more personalized, treatment strategies.


Assuntos
Cisplatino/uso terapêutico , Resistencia a Medicamentos Antineoplásicos/genética , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Neoplasias Ovarianas/tratamento farmacológico , Proteína Grupo D do Xeroderma Pigmentoso/genética , Antineoplásicos/uso terapêutico , Apoptose/efeitos dos fármacos , Apoptose/genética , Linhagem Celular Tumoral , Feminino , Perfilação da Expressão Gênica/métodos , Humanos , Hibridização In Situ , Estimativa de Kaplan-Meier , Pessoa de Meia-Idade , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/metabolismo , Interferência de RNA , Proteína Grupo D do Xeroderma Pigmentoso/metabolismo
9.
J Biomed Nanotechnol ; 12(9): 1774-81, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29345888

RESUMO

It has been reported that gambogic acid (GA), the main active component of gamboge, could directly inhibit and reduce the expression level of P-gp by promoting protein degradation through post-translational proteasome pathway. In this study, the optimal molar ratio of GA/docetaxel (DTX) that could recover the sensitivity of MCF-7/ADR cells to DTX was firstly investigated. Then GA and DTX were loaded simultaneously in PLGA nanoparticles in terms of the optimal ratio. In vitro cell apoptosis and western-blot assays showed that co-delivery of anticancer drugs resulted in enhanced cell apoptosis through the downregulation of the expression level of P-gp. Interestingly, in vivo pharmacokinetic study demonstrated that GA and DTX are released synchronously in blood from the NPs. Finally, the most effective tumor growth inhibition in the MCF-7/ADR human breast tumor xenograft was observed in the co-delivery nanoparticle formulation group in comparison with saline control, free DTX solution and free DTX/GA solution. Taken together, our study demonstrated that DTX/GA PLGA NPs based combination therapy holds significant potential towards the treatment of multidrug-resistant breast cancer.


Assuntos
Antineoplásicos/uso terapêutico , Neoplasias da Mama/tratamento farmacológico , Nanopartículas/uso terapêutico , Taxoides/uso terapêutico , Xantonas/uso terapêutico , Animais , Antineoplásicos/química , Antineoplásicos/farmacocinética , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Docetaxel , Resistência a Múltiplos Medicamentos , Resistencia a Medicamentos Antineoplásicos , Feminino , Humanos , Células MCF-7 , Masculino , Nanopartículas/química , Ratos , Ratos Sprague-Dawley , Taxoides/química , Taxoides/farmacocinética , Taxoides/farmacologia , Xantonas/química , Xantonas/farmacocinética , Xantonas/farmacologia , Ensaios Antitumorais Modelo de Xenoenxerto
10.
Mol Med Rep ; 12(6): 8021-31, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26497752

RESUMO

Improved insight into the molecular and genetic profile of different types of epithelial ovarian cancer (EOC) is required for understanding the carcinogenesis of EOC and may potentially be exploited by future targeted therapies. The aim of the present study was to identify a unique microRNA (miRNA) patterns and key miRNAs, which may assist in predicting progression and prognosis in high­grade serous carcinoma (HGSC) and clear cell carcinoma (CCC). To identify unique miRNA patterns associated with HGSC and CCC, a miRNA microarray was performed using Chinese tumor bank specimens of patients with HGSC or CCC in a retrospective analysis. The expression levels of four deregulated miRNAs were further validated using reverse transcription­quantitative polymerase chain reaction (RT­qPCR) in an external cohort of 42 cases of HGSC and 36 cases of CCC. Kaplan­Meier analysis was performed to analyze the correlation between the expression levels of the four miRNAs and patient prognosis. Among these validated miRNAs, miR­510 was further examined in another cohort of normal ovarian tissues, as well as the HGSC, low­grade serous carcinoma (LGSC) and CCC specimens using RT­qPCR and in situ hybridization. The results revealed that, of the 768 miRNAs analyzed in the microarray, 33 and 50 miRNAs were significantly upregulated and downregulated, respectively, with at least a 2­fold difference in HGSC, compared with CCC. The quantitative analysis demonstrated that miR­510 and miR­129­3p were significantly downregulated, and that miR­483­5p and miR­miR­449a were significantly upregulated in CCC, compared with HGSC (P<0.05), which was consistent with the microarray results. Kaplan­Meier analysis revealed low expression levels of miR­510 and low expression levels of miR­129­3p, advanced International Federation of Gynecology and Obstetrics (FIGO) stage, lymphatic metastasis and that HGSC was significantly associated with the poorer overall survival rates (P<0.05). The expression of miR­510 was significantly higher in the LGSC and CCC tissues, compared with the HGSC and normal ovarian tissues. The results of the present study suggested that different subtypes of EOC have specific miRNA signatures, and that miR­510 may be involved differently in HGSC and CCC. Thus, miR­510 and miR­129­3p may be considered as potential novel candidate clinical biomarkers for predicting the outcome of EOC.


Assuntos
Carcinoma/patologia , Neoplasias Ovarianas/patologia , Adulto , Biomarcadores Tumorais/genética , Carcinoma/genética , Carcinoma/mortalidade , Análise por Conglomerados , Regulação para Baixo , Feminino , Humanos , Estimativa de Kaplan-Meier , Metástase Linfática , MicroRNAs/genética , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Gradação de Tumores , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/mortalidade , Estudos Retrospectivos , Regulação para Cima
11.
Oncol Rep ; 33(2): 591-8, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25482209

RESUMO

MicroRNAs (miRNAs) are involved in regulating the response of cancer cells to various therapeutic interventions, yet their involvement in the chemoresistance of human epithelial ovarian cancer is not fully understood. We found that miR-136 was significantly downregulated in specimens from patients with chemoresistant epithelial ovarian cancer. In the present study, we aimed to clarify the role of miR-136 in regulating the chemoresistance of ovarian cancer. Thirty-four tumor bank specimens and 2 well-established human ovarian cancer cell lines, C13 and OV2008, were used. We found that miR-136 expression was significantly reduced in primary platinum-resistant patients and the ovarian cancer OVC cell line. Enforced expression of miR-136 decreased the chemoresistance to cisplatin in OVC cells through inhibition of cell survival. In addition, we found no association between miR-136 and migration or invasion potential in the ovarian cancer cell lines. However, in the platinum-resistant C13 cell line, the overexpression of miR-136 markedly promoted an apoptotic response to cisplatin. Furthermore, the levels of adducts corrected with their extent of DNA damage/repair, in terms of the percentage of DNA in comet tails, tail length, tail moment (TM), and olive tail moment (OTM), revealed that miR-136 is essential for the repair of cisplatin-induced DNA damage. Our findings suggest that miR-136 may function as an anti-oncogene and deficiency of miR-136 expression in ovarian cancer can induce chemoresistance at least in part by downregulating apoptosis and promoting the repair of cisplatin-induced DNA damage. Thus, miR-136 may provide a biomarker for predicting the chemosensitivity to cisplatin in patients with epithelial ovarian cancer.


Assuntos
Antineoplásicos/administração & dosagem , Cisplatino/administração & dosagem , Resistencia a Medicamentos Antineoplásicos , MicroRNAs/genética , Neoplasias Epiteliais e Glandulares/tratamento farmacológico , Neoplasias Ovarianas/tratamento farmacológico , Carcinoma Epitelial do Ovário , Linhagem Celular Tumoral , Movimento Celular , Dano ao DNA/efeitos dos fármacos , Reparo do DNA , Regulação para Baixo , Feminino , Humanos , Pessoa de Meia-Idade , Invasividade Neoplásica , Neoplasias Epiteliais e Glandulares/genética , Neoplasias Ovarianas/genética
12.
Tumour Biol ; 34(6): 3501-8, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23836287

RESUMO

One of the best prognostic predictors for patients with epithelial ovarian cancer is the Federation of Obstetrics and Gynecology (FIGO) stage at diagnosis. Advanced-stage ovarian serous carcinoma (OSC) generally have poor prognosis. The goal of this study is to develop and validate a miRNA expression profile that can differentiate the OSC at early and advanced stages and study its correlation with the prognosis of OSC. To identify a unique microRNA (miRNA) pattern associated with the progression of OSC at early and advanced stages, a miRNA microarray was performed using Chinese tumor bank specimens of patients with OSC stage I or III in a retrospective analysis. The expression of four dysregulated miRNAs was validated using quantitative real-time polymerase chain reaction (qRT-PCR) in an external cohort of 51 cases of OSC samples at stages I and III. Kaplan-Meier analysis was performed to analyze the correlation between the expression of some miRNAs and prognosis. Of the 768 miRNAs analyzed in the microarray, 26 miRNAs were significantly either up- or downregulated, with at least a 2-fold difference, in OSC stage I compared with stage III. The qRT-PCR results showed that miR-510, miR-509-5p, and miR-508-3p were significantly downregulated and that miR-483-5p was upregulated in stage III OSC compared with stage I, which was consistent with the microarray results. Kaplan-Meier analysis showed low miR-510 expression, low miR-509-5p expression, and advanced FIGO stage, and chemotherapy resistance were significantly associated with poorer overall survival (P < 0.05). Our results suggest that miRNAs may play a role in the progression of OSC, and miR-510 and miR-509-5p may be considered novel-candidate clinical biomarkers for predicting OSC outcome.


Assuntos
Cistadenocarcinoma Seroso/genética , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Neoplasias Ovarianas/genética , Transcriptoma , Adulto , Idoso , Análise por Conglomerados , Cistadenocarcinoma Seroso/patologia , Feminino , Humanos , Estimativa de Kaplan-Meier , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Análise de Sequência com Séries de Oligonucleotídeos , Neoplasias Ovarianas/patologia , Prognóstico , Reação em Cadeia da Polimerase em Tempo Real
13.
Int J Oncol ; 43(3): 839-49, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23787480

RESUMO

The long-term survival for elderly patients with advanced ovarian papillary serous carcinoma (OPSC) does not exceed 30%, and the incidence and prognosis rise continuously after menopause. The aim of this study was to identify the differences in key miRNAs and their potential regulators through miRNA microarray analysis, functional target prediction, and clinical outcome between the elderly patients with advanced OPSC and ovarian clear cell carcinoma (OCC) who all suffered poor prognosis, to identify the pathogenetic basis, and to improve the understanding of the molecular basis of advanced OPCS in elderly patients. Through microarray analysis, we found 52 unique miRNAs with significant fold­change in expression levels, of which 9 were upregulated, whereas 43 were downregulated in OCC patients compared to elderly OPSC patients with advanced stage. Among these prediction miRNAs, miR-30a, miR-30e and miR-505 were found to have some common cancer-related targets. Lower expression of these three miRNAs of advanced OPSC in elderly patients, all associated with significantly poorer survival rate, strongly suggesting that they could be critical oncogenes and take important roles in OPSC etiology in elderly patients with advantaged stage. Functional analyses support the above hypothesis. Their targets, ATF3, STMN1 and MYC suggest that OPSC also has aggressive biological behavior when presented with advanced stage, and support the epidemiology results that incidence and mortality of advanced OPSC increases continuously. miR-30a, miR-30e and miR-505 may be important pathogenetic factors for OPSC in elderly patients with advanced stage. Age could be regarded as a continuous covariate in this process. This may improve the understanding of molecular underpinnings of advanced OPSC in elderly patients, and provide improved diagnostic, prognostic and therapeutic approaches.


Assuntos
Biomarcadores Tumorais/genética , Cistadenocarcinoma Seroso/genética , MicroRNAs/genética , Neoplasias Epiteliais e Glandulares/genética , Neoplasias Ovarianas/genética , Adulto , Fatores Etários , Idoso , Biomarcadores Tumorais/biossíntese , Carcinoma Epitelial do Ovário , Cistadenocarcinoma Seroso/patologia , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , MicroRNAs/biossíntese , Análise em Microsséries , Pessoa de Meia-Idade , Neoplasias Epiteliais e Glandulares/patologia , Neoplasias Ovarianas/patologia , Prognóstico , Taxa de Sobrevida
14.
Methods ; 58(2): 156-63, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22868004

RESUMO

After discovering new miRNAs, it is often difficult to determine their targets and effects on downstream protein expression. In situ hybridization (ISH) and immunohistochemistry (IHC) are two commonly used methods for clinical diagnosis and basic research. We used an optimized technique that simultaneously detects miRNAs, their binding targets and corresponding proteins on transferred serial formalin fixed paraffin embedded (FFPE) sections from patients. Combined with bioinformatics, this method was used to validate the reciprocal expression of specific miRNAs and targets that were detected by ISH, as well as the expression of downstream proteins that were detected by IHC. A complete analysis was performed using a limited number of transferred serial FFPE sections that had been stored for 1-4 years at room temperature. Some sections had even been previously stained with H&E. We identified a miRNA that regulates epithelial ovarian cancer, along with its candidate target and related downstream protein. These findings were directly validated using sub-cellular components obtained from the same patient sample. In addition, the expression of Nephrin (a podocyte marker) and Stmn1 (a recently identified marker related to glomerular development) were confirmed in transferred FFPE sections of mouse kidney. This procedure may be adapted for clinical diagnosis and basic research, providing a qualitative and efficient method to dissect the detailed spatial expression patterns of miRNA pathways in FFPE tissue, especially in cases where only a small biopsy sample can be obtained.


Assuntos
MicroRNAs , Neoplasias Epiteliais e Glandulares , Neoplasias Ovarianas , Proteínas , Adulto , Animais , Carcinoma Epitelial do Ovário , Feminino , Humanos , Imuno-Histoquímica/métodos , Hibridização In Situ/métodos , Proteínas de Membrana/isolamento & purificação , Proteínas de Membrana/metabolismo , Camundongos , MicroRNAs/genética , MicroRNAs/isolamento & purificação , Neoplasias Epiteliais e Glandulares/genética , Neoplasias Epiteliais e Glandulares/metabolismo , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/metabolismo , Inclusão em Parafina , Proteínas/genética , Proteínas/isolamento & purificação , Estatmina/isolamento & purificação , Estatmina/metabolismo
15.
J Cell Sci ; 124(Pt 3): 359-68, 2011 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-21224400

RESUMO

MicroRNAs (miRNAs) are small noncoding RNAs that have important roles in gene regulation. We have previously reported that activin receptor-like kinase 7 (ALK7) and its ligand, Nodal, induce apoptosis in human epithelial ovarian cancer cells. In this study, we examined the regulation of ALK7 by miRNAs and demonstrate that miR-376c targets ALK7. Ectopic expression of miR-376c significantly increased cell proliferation and survival, enhanced spheroid formation and blocked Nodal-induced apoptosis. Interestingly, overexpression of miR-376c blocked cisplatin-induced cell death, whereas anti-miR-376c enhanced the effect of cisplatin. These effects of miR-376c were partially compensated by the overexpression of ALK7. Moreover, in serous carcinoma samples taken from ovarian cancer patients who responded well to chemotherapy, strong ALK7 staining and low miR-376c expression was detected. By contrast, ALK7 expression was weak and miR-376c levels were high in samples from patients who responded poorly to chemotherapy. Finally, treatment with cisplatin led to an increase in expression of mRNA encoding Nodal and ALK7 but a decrease in miR-376c levels. Taken together, these results demonstrate that the Nodal-ALK7 pathway is involved in cisplatin-induced cell death in ovarian cancer cells and that miR-376c enhances proliferation, survival and chemoresistance by targeting, at least in part, ALK7.


Assuntos
Receptores de Ativinas Tipo I/metabolismo , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/genética , Resistencia a Medicamentos Antineoplásicos/genética , MicroRNAs , Receptores de Ativinas Tipo I/efeitos dos fármacos , Receptores de Ativinas Tipo I/genética , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Apoptose/genética , Carcinoma Epitelial do Ovário , Linhagem Celular Tumoral , Cisplatino/farmacologia , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , Neoplasias Epiteliais e Glandulares/genética , Neoplasias Epiteliais e Glandulares/metabolismo , Neoplasias Epiteliais e Glandulares/patologia , Proteína Nodal/efeitos dos fármacos , Proteína Nodal/genética , Proteína Nodal/metabolismo , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/metabolismo , Neoplasias Ovarianas/patologia
16.
Mol Cell Biochem ; 304(1-2): 135-42, 2007 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-17607508

RESUMO

Previous studies showed that nitricoxide synthase (NOS) and oxidative stress can induce skeletal muscle atrophy in the muscular dystrophy and inclusion-body myopathy. There is a correlation between NOS and oxidative stress. However, it is not clear, whether there are some changes of the NOS activity in prolonged alcoholic myopathy (PAM), and whether NOS activity has relation to amyotrophy of PAM. We established experimental alcoholic myopathy model of rats by prolonged alcohol intake. We found that there is a reduction in GSH-px (P < 0.05) and an increase of SOD (P < 0.05), MDA (P < 0.05) and iNOS (P < 0.05) in the plantaris of the experimental group by spectrophotometer. In the soleus of the experimental group, except for MDA showed an increase (P < 0.05), the other enzymes showed no obvious difference (P > 0.05). The immunohistochemistry results showed that there was obvious expression of iNOS in the cytoplasm of plantaris in the experimental group and there was no expression of iNOS in the control group. There was a decrease of nNOS expression on the membranes of the plantaris cells in the experimental group by immunofluorescence. Meanwhile, we found the expression of nNOS in some cytoplasm. Our results suggested that NOS might be an important factor during the development of PAM. We could infer that there are some disturbances with regard to output and scavenging of free radical in PAM. Alcohol can induce the oxidative stress reaction and further result in imbalance of the oxidant-antioxidant status in the organism.


Assuntos
Consumo de Bebidas Alcoólicas/efeitos adversos , Doenças Musculares/induzido quimicamente , Doenças Musculares/enzimologia , Óxido Nítrico Sintase/fisiologia , Estresse Oxidativo , Consumo de Bebidas Alcoólicas/patologia , Animais , Peso Corporal/efeitos dos fármacos , Doença Crônica , Modelos Animais de Doenças , Masculino , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/patologia , Músculo Esquelético/ultraestrutura , Doenças Musculares/metabolismo , Doenças Musculares/patologia , Óxido Nítrico Sintase/genética , Óxido Nítrico Sintase/metabolismo , Tamanho do Órgão/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley
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