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1.
Ital J Pediatr ; 45(1): 170, 2019 Dec 27.
Artigo em Inglês | MEDLINE | ID: mdl-31881905

RESUMO

Radiation therapy represents an important approach in the therapeutic management of children and adolescents with malignant tumors and its application with modern techniques - including Proton Beam Therapy (PBT) - is of great interest. In particular, potential radiation-induced injuries and secondary malignancies - also associated to the prolonged life expectancy of patients - are still questions of concern that increase the debate on the usefulness of PBT in pediatric treatments. This paper presents a literary review of current applications of PBT in non-Central Nervous System pediatric tumors (such as retinoblastoma, Hodgkin Lymphoma, Wilms tumor, bone and soft tissues sarcomas). We specifically reported clinical results achieved with PBT and dosimetric comparisons between PBT and the most common photon-therapy techniques. The analysis emphasizes that PBT minimizes radiation doses to healthy growing organs, suggesting for reduced risks of late side-effects and radiation-induced secondary malignancies. Extended follow up and confirms by prospective clinical trials should support the effectiveness and long-term tolerance of PBT in the considered setting.


Assuntos
Neoplasias/radioterapia , Terapia com Prótons , Neoplasias Ósseas/radioterapia , Doença de Hodgkin/radioterapia , Humanos , Órgãos em Risco , Doses de Radiação , Dosagem Radioterapêutica , Retinoblastoma/radioterapia , Sarcoma/radioterapia , Tumor de Wilms/radioterapia
4.
Pathol Res Pract ; 215(12): 152641, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31727502

RESUMO

BACKGROUND: Retinoblastoma (RB) is the most common primary intraocular malignancy in children. Accumulating evidences have clarified that microRNAs (miRNAs) modulated signaling molecules by acting as oncogenes or tumor-suppressor genes in RB. Thus, in our study, we aimed to investigate the function of miR-129-5p in RB cells through PI3K/AKT signaling pathway by targeting PAX6. Two RB cell lines, Y79 and WERI-Rb-1, were selected in our study, followed by transfection of miR-129-5p inhibitor or si-PAX6 to explore the regulatory role of miR-129-5p in RB cell proliferation, invasion and migration. MATERIAL AND METHODS: Dual-luciferase assay was used for the detection of targeting relationship between miR-129-5p and PAX6. Besides, western blot analysis was applied to detect expression of cell cycle-related factors (CDK2 and Cyclin E) and PI3K/AKT signaling pathway-related factors (p-AKT and AKT). Nude mice tumorigenesis experiment was used to evaluate the effect of miR-129a-5p on RB growth in vivo. RESULTS: miR-129-5p was down-regulated in RB cell lines. miR-129-5p directly targeted the 3'-untranslated region of PAX6. Artificial down-regulation of miR-129-5p promoted cell proliferation, migration and invasion in RB cell lines Y79 and WERI-Rb-1, and promoted RB growth in vivo via PI3K/AKT signaling pathway, which could be reversed by transfection with silencing PAX6. CONCLUSION: This study provides evidences that RB progression was suppressed by overexpressed miR-129-5p via direct targeting of PAX6 through PI3K/AKT signaling pathway, which may provide a molecular basis for better treatment for RB.


Assuntos
Movimento Celular , Proliferação de Células , MicroRNAs/metabolismo , Fator de Transcrição PAX6/metabolismo , Fosfatidilinositol 3-Quinase/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Neoplasias da Retina/enzimologia , Retinoblastoma/enzimologia , Animais , Ciclo Celular , Linhagem Celular Tumoral , Regulação Neoplásica da Expressão Gênica , Humanos , Camundongos Endogâmicos BALB C , Camundongos Nus , MicroRNAs/genética , Invasividade Neoplásica , Fator de Transcrição PAX6/genética , Neoplasias da Retina/genética , Neoplasias da Retina/patologia , Retinoblastoma/genética , Retinoblastoma/patologia , Transdução de Sinais , Carga Tumoral
5.
Genes (Basel) ; 10(11)2019 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-31683923

RESUMO

The RB1 gene is the first described human tumor suppressor gene and plays an integral role in the development of retinoblastoma, a pediatric malignancy of the eye. Since its discovery, the stepwise characterization and cloning of RB1 have laid the foundation for numerous advances in the understanding of tumor suppressor genes, retinoblastoma tumorigenesis, and inheritance. Knowledge of RB1 led to a paradigm shift in the field of cancer genetics, including widespread acceptance of the concept of tumor suppressor genes, and has provided crucial diagnostic and prognostic information through genetic testing for patients affected by retinoblastoma. This article reviews the long history of RB1 gene research, characterization, and cloning, and also discusses recent advances in retinoblastoma genetics that have grown out of this foundational work.


Assuntos
Biomarcadores Tumorais/genética , Genes Supressores de Tumor , Proteínas de Ligação a Retinoblastoma/genética , Retinoblastoma/genética , Ubiquitina-Proteína Ligases/genética , Biomarcadores Tumorais/metabolismo , Clonagem Molecular , Humanos , Retinoblastoma/diagnóstico , Proteínas de Ligação a Retinoblastoma/metabolismo , Ubiquitina-Proteína Ligases/metabolismo
6.
BMJ Case Rep ; 12(10)2019 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-31653630

RESUMO

Retinoblastoma is the most common intraocular malignancy of infancy which frequently manifests with a white pupillary reflex. We report a case of delayed presentation of a child with retinoblastoma in his left eye because parents thought the change in iris colour in this eye was due to the innocent heterochromia irides that was previously diagnosed in his elder sibling. This late presentation necessitated enucleation of the affected eye followed by chemotherapy.


Assuntos
Neoplasias da Retina/cirurgia , Retinoblastoma/cirurgia , Diagnóstico Diferencial , Enucleação Ocular , Humanos , Lactente , Doenças da Íris/diagnóstico , Masculino , Transtornos da Pigmentação/diagnóstico
7.
Klin Onkol ; 32(5): 375-379, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31610671

RESUMO

BACKGROUND: The MTHFR 677C>T, 1298A>C and MTR 2756A>G polymorphisms have been investigated in several different cancer types. However, the role of these polymorphisms in the development of retinoblastoma remains unclear. Here, we have evaluated the association of the MTHFR 677C>T, 1298A>C and MTR 2756A>G polymorphisms with the risk of retinoblastoma in Iranian children. METHODS: The MTHFR 677C>T, 1298A>C and MTR 2756A>G polymorphisms in 66 patients with retinoblastoma and 99 age-and gender-matched healthy controls were detected on the ABI PRISMs 7500 Real-Time PCR System. The association between these polymorphisms and the risk of retinoblastoma was analysed by an odds ratio with a 95% confidence interval. RESULTS: Our results showed a significant association between the MTR 2756A>G polymorphism and the risk of retinoblastoma. In the MTR 2756A>G polymorphism, the AG (39.4%) and GG (9.1%) genotype frequencies in the cases were found to be higher in comparison with the controls, showing a significant difference (p < 0.05). However, no significant difference was observed in the allelic or genotypic frequencies for both the MTHFR 677C>T and 1298A>C polymorphisms in the retinoblastoma patients of the controls (p > 0.05). CONCLUSIONS: Our results suggested that the MTR 2756A>G polymorphism might be associated with an increased risk of retinoblastoma in Iranian children. However, the results show that the MTHFR 677C>T and 1298A>C polymorphisms are not significantly associated with an increased risk of retinoblastoma.


Assuntos
Predisposição Genética para Doença , Metilenotetra-Hidrofolato Redutase (NADPH2)/genética , Neoplasias da Retina/genética , Retinoblastoma/genética , Estudos de Casos e Controles , Criança , Pré-Escolar , Feminino , Genótipo , Humanos , Lactente , Recém-Nascido , Irã (Geográfico)/epidemiologia , Masculino , Polimorfismo de Nucleotídeo Único , Fatores de Risco
8.
BMJ Case Rep ; 12(10)2019 Oct 13.
Artigo em Inglês | MEDLINE | ID: mdl-31611224

RESUMO

We report a case of a man aged 35 years who presented with the chief complaint of painless diminution of vision in the right eye for 4 months. Examination revealed a large inferior retinochoroidal mass along with retinal detachment. An anterior choroidal mass with moderate internal reflectivity was seen on B-scan ocular ultrasonography and MRI and CT scan were indicative of a mitotic aetiology. Fluorodeoxyglucose-positron emission tomography scan ruled out any other systemic foci of involvement. Based on the above findings, a provisional diagnosis of amelanotic choroidal melanoma was made and he was taken up for choroidal aspiration biopsy, wherein the cytopathology report revealed hypercellularity with no identifiable pigments. In view of the above, a diagnosticandtherapeutic enucleation was performed; however, the histopathology report of the enucleated specimen revealed poorly differentiated retinoblastoma. This case highlights that the diagnosis of retinoblastoma should be kept in mind even in adult patients.


Assuntos
Neoplasias da Coroide/diagnóstico , Melanoma Amelanótico/diagnóstico , Neoplasias da Retina/diagnóstico , Retinoblastoma/diagnóstico , Adulto , Diagnóstico Diferencial , Humanos , Masculino
9.
Int J Biol Macromol ; 141: 997-1003, 2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31521654

RESUMO

Melphalan (MEL) is an effective chemotherapeutic agent for treatment of retinoblastoma (Rb) which is the most common childhood malignancy. However, the inherent cardiopulmonary toxicity and hazardous integration limit its therapeutic effect on RB. N-Acetylheparosan (AH), a natural heparin-like polysaccharide in mammals with long circulation effect and good biocompatibility, was linked by d-α-tocopherol acid succinate (VES) via and cystamine (CYS) to synthesize reduction-responsive N-acetylheparosan-CYS-Vitamin E succinate (AHV) copolymers. In addition, CYS was replaced by adipic acid dihydrazide (ADH) to obtain a control of non-reduction-responsive polymers N-acetylheparosan-ADH-Vitamin E succinate (ADV). MEL-loaded AHV micelles (MEL/AHV) as well as ADV micelles (MEL/ADV) were prepared with small particle size and high drug loading content. In vitro drug release showed that MEL/AHV micelles presented obvious reduction-triggered release behavior compared with MEL/ADV. In vitro antitumor effects were investigated using WERI-Rb-1 retinoblastoma cells. Cytotoxicity experiments showed that the IC50 of MEL/AHV was significantly lower than that of free MEL and MEL/ADV, suggesting that MEL/AHV enhanced the cytotoxicity against retinoblastoma cells. Furthermore, MEL/AHV micelles were more easily uptaken by multiple pathways compared with MEL/ADV and free MEL. Therefore, MEL/AHV might be a potential delivery system for enhanced delivery of melphalan to Rb cells.


Assuntos
Antineoplásicos Alquilantes/administração & dosagem , Antineoplásicos Alquilantes/química , Melfalan/administração & dosagem , Melfalan/química , Micelas , Antineoplásicos Alquilantes/síntese química , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Liberação Controlada de Fármacos , Humanos , Espectroscopia de Ressonância Magnética , Melfalan/síntese química , Modelos Biológicos , Polímeros/química , Retinoblastoma
10.
Middle East Afr J Ophthalmol ; 26(2): 107-109, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31543669

RESUMO

Trauma is the major reason for globe loss in adults; however, there are less data regarding the causes for globe loss in children. We reviewed the underlying diagnoses of children who underwent ocular prosthesis fitting over a 1-year period at a referral eye hospital in the Middle East and found retinoblastoma, trauma, and congenital microphthalmia or anophthalmia to be the most common diagnoses, respectively. Enucleation and evisceration were the most common procedures and were exclusively performed for retinoblastoma and trauma, respectively. Ocular morbidity from the most common diagnoses related to pediatric globe loss in the region could be decreased by improved family education, safety precautions, and genetic counseling.


Assuntos
Anoftalmia/complicações , Olho Artificial , Microftalmia/complicações , Implantes Orbitários , Ajuste de Prótese , Retinoblastoma/complicações , Anoftalmia/diagnóstico , Anoftalmia/cirurgia , Criança , Pré-Escolar , Enucleação Ocular , Evisceração do Olho , Traumatismos Oculares/cirurgia , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Microftalmia/diagnóstico , Microftalmia/cirurgia , Oriente Médio , Implantação de Prótese , Encaminhamento e Consulta , Retinoblastoma/diagnóstico , Retinoblastoma/cirurgia , Estudos Retrospectivos
12.
Med Sci Monit ; 25: 6639-6648, 2019 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-31483776

RESUMO

BACKGROUND Biglycan (BGN) is an extracellular matrix (ECM) protein that regulates the growth of epithelial cells. The mammalian target of rapamycin (mTOR) inhibitor, rapamycin, is a treatment for advanced retinoblastoma. This study aimed to investigate the effects of expression of BGN on the response of human WERI-Rb-1 retinoblastoma cells to rapamycin and to investigate the associated signaling pathways. MATERIAL AND METHODS BGN gene expression was induced in human WERI-Rb-1 retinoblastoma cells, which were incubated with rapamycin at doses of 0, 5, 10, 20, 30, and 50 µg/ml. Cells were treated with the PI3K/Akt pathway inhibitor, LY294002. The MTT assay determined the rate of cell inhibition. Real-time polymerase chain reaction (RT-PCR) was performed to measure BGN gene expression using RT²-PCR. Western blot detected the protein levels of BGN, p-PI3K, p-Akt, nuclear NF-kappaB, and p65. RESULTS Rapamycin impaired cell growth, induced cell apoptosis, and suppressed the expression levels of p-PI3K, p-Akt, nuclear NF-kappaB, and p65. Overexpression of the BGN gene restored growth potential and inhibited apoptosis and was associated with the activation of the PI3K/Akt-mediated NF-kappaB pathway. In cells that overexpressed BGN, inhibition of the PI3K/Akt pathway by LY294002 increased the sensitivity of human WERI-Rb-1 retinoblastoma cells to rapamycin. CONCLUSIONS Overexpression of BGN induced rapamycin resistance in WERI-Rb-1 retinoblastoma cells by activating PI3K/Akt/NF-kappaB signaling.


Assuntos
Biglicano/metabolismo , Resistencia a Medicamentos Antineoplásicos , NF-kappa B/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Retinoblastoma/metabolismo , Retinoblastoma/patologia , Sirolimo/farmacologia , Apoptose/efeitos dos fármacos , Biglicano/genética , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Concentração Inibidora 50 , Retinoblastoma/genética , Transdução de Sinais
13.
BMC Cancer ; 19(1): 895, 2019 Sep 09.
Artigo em Inglês | MEDLINE | ID: mdl-31500597

RESUMO

BACKGROUND: Chimeric antigen receptor (CAR)-based T cell therapy is in early clinical trials to target the neuroectodermal tumor, neuroblastoma. No preclinical or clinical efficacy data are available for retinoblastoma to date. Whereas unilateral intraocular retinoblastoma is cured by enucleation of the eye, infiltration of the optic nerve indicates potential diffuse scattering and tumor spread leading to a major therapeutic challenge. CAR-T cell therapy could improve the currently limited therapeutic strategies for metastasized retinoblastoma by simultaneously killing both primary tumor and metastasizing malignant cells and by reducing chemotherapy-related late effects. METHODS: CD171 and GD2 expression was flow cytometrically analyzed in 11 retinoblastoma cell lines. CD171 expression and T cell infiltration (CD3+) was immunohistochemically assessed in retrospectively collected primary retinoblastomas. The efficacy of CAR-T cells targeting the CD171 and GD2 tumor-associated antigens was preclinically tested against three antigen-expressing retinoblastoma cell lines. CAR-T cell activation and exhaustion were assessed by cytokine release assays and flow cytometric detection of cell surface markers, and killing ability was assessed in cytotoxic assays. CAR constructs harboring different extracellular spacer lengths (short/long) and intracellular co-stimulatory domains (CD28/4-1BB) were compared to select the most potent constructs. RESULTS: All retinoblastoma cell lines investigated expressed CD171 and GD2. CD171 was expressed in 15/30 primary retinoblastomas. Retinoblastoma cell encounter strongly activated both CD171-specific and GD2-specific CAR-T cells. Targeting either CD171 or GD2 effectively killed all retinoblastoma cell lines examined. Similar activation and killing ability for either target was achieved by all CAR constructs irrespective of the length of the extracellular spacers and the co-stimulatory domain. Cell lines differentially lost tumor antigen expression upon CAR-T cell encounter, with CD171 being completely lost by all tested cell lines and GD2 further down-regulated in cell lines expressing low GD2 levels before CAR-T cell challenge. Alternating the CAR-T cell target in sequential challenges enhanced retinoblastoma cell killing. CONCLUSION: Both CD171 and GD2 are effective targets on human retinoblastoma cell lines, and CAR-T cell therapy is highly effective against retinoblastoma in vitro. Targeting of two different antigens by sequential CAR-T cell applications enhanced tumor cell killing and preempted tumor antigen loss in preclinical testing.


Assuntos
Terapia Baseada em Transplante de Células e Tecidos , Gangliosídeos/imunologia , Molécula L1 de Adesão de Célula Nervosa/imunologia , Receptores de Antígenos Quiméricos , Retinoblastoma/terapia , Linfócitos T/metabolismo , Linhagem Celular Tumoral , Criança , Pré-Escolar , Citotoxicidade Imunológica , Feminino , Humanos , Lactente , Masculino , Retinoblastoma/imunologia , Retinoblastoma/metabolismo , Estudos Retrospectivos , Linfócitos T/imunologia
14.
Pediatr Blood Cancer ; 66(11): e27959, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31423715

RESUMO

Retinoblastoma is an ocular tumor that occurs in young children, in either heritable or sporadic manner. The relative rarity of retinoblastoma, and the need for expensive equipment, anesthesia, and pediatric ophthalmologic expertise, are barriers for effective treatment in developing countries. Also, with an average age-adjusted incidence of two to five cases per million children, patient number limits development of local expertise in countries with small populations. Lebanon is a small country with a population of approximately 4.5 million. In 2012, a comprehensive retinoblastoma program was formalized at the Children's Cancer Institute (CCI) at the American University of Beirut Medical Center, and resources were allocated for efficient interdisciplinary coordination to attract patients from neighboring countries such as Syria and Iraq, where such specialized therapy is also lacking. Through this program, care was coordinated across hospitals and borders such that patients would receive scheduled chemotherapy at their institution, and monthly retinal examinations and focal laser therapy at the CCI in Lebanon. Our results show the feasibility of successful collaboration across borders, with excellent patient and physician adherence to treatment plans. This was accompanied by an increase in patient referrals, which enables continued expertise development. However, the majority of patients presented with advanced intraocular disease, necessitating enucleation in 90% of eyes in unilateral cases, and more than 50% of eyes in bilateral cases. Future efforts need to focus on expanding the program that reaches to additional hospitals in both countries, and promoting early diagnosis, for further improvement of globe salvage rates.


Assuntos
Institutos de Câncer/organização & administração , Países em Desenvolvimento , Hospitais Universitários/organização & administração , Internacionalidade , Colaboração Intersetorial , Neoplasias da Retina/terapia , Retinoblastoma/terapia , Institutos de Câncer/economia , Terapia Combinada/economia , Terapia Combinada/métodos , Diagnóstico Tardio , Gerenciamento Clínico , Estudos de Viabilidade , Feminino , Aconselhamento Genético , Hospitais Universitários/economia , Humanos , Incidência , Lactente , Recém-Nascido , Estimativa de Kaplan-Meier , Líbano/epidemiologia , Masculino , Oriente Médio/epidemiologia , Equipe de Assistência ao Paciente , Encaminhamento e Consulta , Neoplasias da Retina/diagnóstico , Neoplasias da Retina/epidemiologia , Retinoblastoma/diagnóstico , Retinoblastoma/economia , Retinoblastoma/epidemiologia , Resultado do Tratamento , Estados Unidos
15.
Stem Cell Res ; 39: 101517, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31404748

RESUMO

Biallelic inactivation of the retinoblastoma tumor suppressor gene (RB1) causes formation of retinoblastoma, a retinal eye tumor occurring in early childhood. Using the CRISPR/Cas9 nickase system, exon 1 of RB1 was deleted, including the RB1 promoter. As a result, sublines were generated carrying deletions of RB1 exon 1/promoter on one or both alleles.


Assuntos
Éxons/genética , Regiões Promotoras Genéticas/genética , Retinoblastoma/genética , Alelos , Sistemas CRISPR-Cas/genética , Imunofluorescência , Heterozigoto , Humanos , Cariotipagem , Masculino , Repetições de Microssatélites/genética , Proteínas de Ligação a Retinoblastoma/genética , Ubiquitina-Proteína Ligases/genética
16.
Int J Mol Sci ; 20(17)2019 Aug 24.
Artigo em Inglês | MEDLINE | ID: mdl-31450568

RESUMO

Trefoil factor family peptide 3 (TFF3) is supposed to have tumor suppressive functions in retinoblastoma (RB), but the functional pathway is not completely understood. In the study presented, we investigated the downstream pathway of TFF3 signaling in Y79 RB cells. Results from pG13-luciferase reporter assays and western blot analyses indicate induced p53 activity with an upregulation of miR-34a after TFF3 overexpression. Expression levels of the predicted miR-34a target epithelial membrane protein 1 (EMP1) are reduced after TFF3 overexpression. As revealed by WST-1 assay, BrdU, and DAPI cell counts viability and proliferation of Y79 cells significantly decrease following EMP1 knockdown, while apoptosis levels significantly increase. Opposite effects on Y79 cells' growth could be shown after EMP1 overexpression. Caspase assays showed that EMP1 induced apoptosis after overexpression is at least partially caspase-3/7 dependent. Colony formation and soft agarose assays, testing for anchorage independent growth, revealed that EMP1 overexpressing Y79 cells have a significantly higher ability to form colonies. In in ovo chicken chorioallantoic membrane (CAM) assays inoculated EMP1 overexpressing Y79 cells form significantly larger CAM tumors. Moreover, miR-34a overexpression increases sensitivity of Y79 cells towards RB chemotherapeutics, however, without involvement of EMP1. In summary, the TFF3 signaling pathway in Y79 RB cells involves the activation of p53 with downstream induction of miR-34a and subsequent inhibition of EMP1.


Assuntos
Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Proteínas de Neoplasias/genética , Interferência de RNA , Receptores de Superfície Celular/genética , Transdução de Sinais , Proteína Supressora de Tumor p53/metabolismo , Apoptose/genética , Linhagem Celular Tumoral , Resistencia a Medicamentos Antineoplásicos/genética , Técnicas de Silenciamento de Genes , Genes Reporter , Humanos , Proteínas de Neoplasias/metabolismo , Receptores de Superfície Celular/metabolismo , Retinoblastoma/genética , Retinoblastoma/metabolismo , Fator Trefoil-3
17.
Mol Med Rep ; 20(4): 3440-3447, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31432120

RESUMO

Abnormal expression of microRNAs (miRNAs/miRs) has been previously reported in various types of human cancer, such as retinoblastoma (RB). Dysregulated miRNAs have been demonstrated to be important epigenetic regulators of numerous biological events associated with RB. Therefore, improved understanding of the precise roles of miRNAs in RB is required to develop novel therapeutic strategies for the treatment of patients with this disease. In the present study, reverse transcription­quantitative polymerase chain reaction (RT­qPCR) was performed to detect miR­330 expression in RB tissues and cell lines. The effects of miR­330 overexpression on the viability and invasion of RB cells were determined using MTT and Matrigel®­based invasion assays, respectively. The mechanisms underlying the activity of miR­330 in RB cells were investigated via bioinformatics analysis, luciferase reporter assays, and RT­qPCR and western blot analyses. It was revealed that the levels of miR­330 expression were significantly downregulated in RB tissues and cell lines compared with in control healthy tissues and cells, respectively. Overexpression of miR­330 in RB cells significantly reduced the viability and invasion of cells in vitro. Additionally, ρ­associated coiled­coil containing protein kinase 1 (ROCK1) was identified as a putative target of miR­330 using bioinformatics analysis. Subsequent experiments revealed that miR­330 interacted with the 3'­untranslated region of ROCK1 and downregulated its expression in RB cells. Furthermore, the expression levels of ROCK1 were increased in RB tissues compared with healthy controls and negatively correlated with miR­330 expression. Finally, upregulation of ROCK1 expression reversed the miR­330­induced inhibition of the viability and invasion of RB cells. Collectively, these results suggested that miR­330 exhibits tumor­suppressor activity in the development of RB by directly targeting ROCK1, indicating that restoration of miR­330 expression may be a promising therapeutic technique in the treatment of patients with RB.


Assuntos
Regulação para Baixo , Regulação Neoplásica da Expressão Gênica , MicroRNAs/biossíntese , Proteínas de Neoplasias/biossíntese , RNA Neoplásico/biossíntese , Retinoblastoma/metabolismo , Quinases Associadas a rho/biossíntese , Adolescente , Adulto , Linhagem Celular Tumoral , Sobrevivência Celular , Feminino , Humanos , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , Invasividade Neoplásica , Proteínas de Neoplasias/genética , RNA Neoplásico/genética , Retinoblastoma/genética , Retinoblastoma/patologia , Quinases Associadas a rho/genética
18.
Nano Lett ; 19(9): 6410-6417, 2019 09 11.
Artigo em Inglês | MEDLINE | ID: mdl-31442373

RESUMO

Gene therapy is promising for chronic posterior ocular diseases, which are causal factors for severe vision impairment and even blindness worldwide. However, the inherent absorption barriers of the eye restrict intraocular delivery of therapeutic nucleic acids via topical instillation. Safe and efficient nonviral vectors for ocular gene therapy are still unmet clinical desires. Herein, an octopus-like flexible multivalent penetratin (MVP) was designed to facilitate condensation and delivery of therapeutic nucleic acids using multiarm polyethylene glycol (PEG) as a core and conjugating penetratin at each end of the PEG arms as outspread tentacles. Among the MVPs, 8-valent penetratin (8VP) stably compacted nucleic acids into positively charged polyplexes smaller than 100 nm, promoting cellular uptake efficiency (approaching 100%) and transfection rate (over 75%). After being instilled into the conjunctival sac, 8VP enabled rapid (<10 min) and prolonged (>6 h) distribution of nucleic acids in the retina via a noncorneal pathway. In a retinoblastoma-bearing mice model, topical instillation of 8VP/siRNA efficiently inhibited the protein expression of intraocular tumor without toxicity. MVP is advantageous over the commercial transfection reagent in safety and efficiency, and therefore provides a promising vector for noninvasive intraocular gene delivery.


Assuntos
Peptídeos Penetradores de Células , Túnica Conjuntiva/metabolismo , Neoplasias Oculares , Terapia Genética , RNA Interferente Pequeno , Retinoblastoma , Transfecção , Animais , Linhagem Celular Tumoral , Peptídeos Penetradores de Células/química , Peptídeos Penetradores de Células/farmacocinética , Peptídeos Penetradores de Células/farmacologia , Túnica Conjuntiva/patologia , Neoplasias Oculares/genética , Neoplasias Oculares/metabolismo , Neoplasias Oculares/patologia , Neoplasias Oculares/terapia , Humanos , Injeções Intraoculares , Camundongos , RNA Interferente Pequeno/química , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/farmacocinética , RNA Interferente Pequeno/farmacologia , Retinoblastoma/genética , Retinoblastoma/metabolismo , Retinoblastoma/patologia , Retinoblastoma/terapia
19.
Biomed Pharmacother ; 118: 109111, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31336343

RESUMO

Aberrant expression of microRNAs plays an important role in the pathogenesis and progression of retinoblastoma. MiR-25, a member of the miR-106b˜25 cluster, has been reported to be abnormally expressed in retinoblastoma, but the exact role of it remains unclear. In our study, we found that miR-25-3p was upregulated in retinoblastoma tissues and cell lines. Enforced expression of miR-25-3p in retinoblastoma cell line WERI-RB-1 increased cell growth, colony formation, anchorage-independent growth, cell migration and invasion in vitro and tumor xenograft growth in vivo. In contrast, inhibited miR-25-3p expression in retinoblastoma cell line Y79 suppressed cell growth, colony formation, anchorage-independent growth, cell migration and invasion. Through luciferase reporter assay, we found that phosphatase and tensin homolog (PTEN) was a direct target of miR-25-3p. This was verified by western blot that miR-25-3p overexpression suppressed PTEN and activated Akt signaling. In addition, miR-25-3p was found to promote epithelial-mesenchymal transition (EMT) of WERI-RB-1 cells through PTEN/Akt pathway. Western blot analysis revealed that miR-25-3p overexpression increased Vimentin and Snail expression, and suppressed E-cadherin expression, but this could be reversed by restoring PTEN. Moreover, LY294002 treatment or restoring PTEN expression abolished the effects of miR-25-3p on cell invasion, colony formation and anchorage-independent growth in vitro and tumor xenograft growth in vivo. Taken together, our results suggested that miR-25-3p promotes malignant transformation of retinoblastoma cells by suppressing PTEN.


Assuntos
Transição Epitelial-Mesenquimal , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , PTEN Fosfo-Hidrolase/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Neoplasias da Retina/genética , Retinoblastoma/genética , Animais , Linhagem Celular Tumoral , Sobrevivência Celular/genética , Transição Epitelial-Mesenquimal/genética , Células HEK293 , Humanos , Camundongos Nus , PTEN Fosfo-Hidrolase/genética , Fosforilação , Proteínas Proto-Oncogênicas c-akt/genética , Neoplasias da Retina/metabolismo , Neoplasias da Retina/patologia , Retinoblastoma/metabolismo , Retinoblastoma/patologia , Transdução de Sinais , Regulação para Cima , Ensaios Antitumorais Modelo de Xenoenxerto
20.
BMC Cancer ; 19(1): 693, 2019 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-31307410

RESUMO

BACKGROUND: To evaluate the safety and efficacy of intra-arterial chemotherapy (IAC) for the primary or secondary treatment of infants diagnosed with advanced retinoblastoma before 3 months of age. METHODS: This single-center retrospective study included 39 infants (42 eyes) aged ≤3 months who were diagnosed with unilateral or bilateral advanced intraocular retinoblastoma (group D and E eyes) and received IAC as primary or secondary treatment between June 2012 and February 2017. Based on each patient's therapeutic history and response to chemotherapeutic drugs, melphalan, topotecan, and/or carboplatin were used for IAC. The main outcomes included the technical success rate for IAC, survival rates, and adverse events. RESULTS: In total, 29 and 13 eyes received IAC as primary and secondary treatments, respectively. Catheterization was successful in 136 of 137 procedures. All eyes in the secondary IAC group had previously received intravenous chemotherapy. The mean number of IAC sessions for each eye was 3 (range, 2-6). The 2-year ocular survival rates were 80.7% (95% confidence interval [CI], 58.9-91.7) in the primary IAC group and 91.7% (95% CI, 53.9-98.8) in the secondary IAC group. During the follow-up period, 1 patient with unilateral disease (group E) developed extraocular disease and died. The 2-year recurrence-free survival rates in the primary and secondary IAC groups were 71.9% (95% CI, 49.4-85.7) and 75.0% (95% CI, 40.8-91.2), respectively. During each catheterization procedure, the main complications included eyelid erythema (2.4%), fundus hemorrhage (11.9%), myelosuppression (7.7%), transient vomiting and hair loss (2.6%), and transient pancytopenia (2.6%). Prolonged complications included phthisis bulbi (19.0%), vision loss (19.0%), poor vision (9.5%), and cataract (2.4%). There was no case of stroke, neurological impairment, secondary malignant tumor, or metastasis. CONCLUSIONS: Our findings suggest that IAC, whether primary or secondary, is effective and fairly safe for the management of advanced retinoblastoma in infants aged < 3 months. However, adverse events related to intra-arterial injection and the visual outcomes cannot be neglected and require further investigation.


Assuntos
Antineoplásicos Fitogênicos/uso terapêutico , Carboplatina/uso terapêutico , Etoposídeo/uso terapêutico , Infusões Intra-Arteriais/efeitos adversos , Neoplasias da Retina/tratamento farmacológico , Retinoblastoma/tratamento farmacológico , Vincristina/uso terapêutico , Antineoplásicos Fitogênicos/efeitos adversos , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Carboplatina/efeitos adversos , Cateterismo/efeitos adversos , Pré-Escolar , Etoposídeo/efeitos adversos , Feminino , Seguimentos , Humanos , Lactente , Recém-Nascido , Masculino , Neoplasias da Retina/mortalidade , Retinoblastoma/mortalidade , Estudos Retrospectivos , Taxa de Sobrevida , Resultado do Tratamento , Vincristina/efeitos adversos
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