RESUMO
BACKGROUND: Long non-coding RNAs (lncRNAs) exert a significant role in carcinogenesis. lncRNA KCNQ1OT1 is detected in many tumors and is considered as an oncogene. The expression and mechanism of KCNQ1OT1 in retinoblastoma (Rb) are not clearly elucidated. METHODS: KCNQ1OT1, miR-134 and TRIM44 mRNA expression were examined by a quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). Proliferation, migration and invasion of Weri-Rb1 and Y79 cells were tested by cell counting kit-8, colony formation, scratch and transwell assays. Meanwhile, the regulatory relationships among KCNQ1OT1, miR-134 and TRIM44 were clarified by several biological experiments, including dual-luciferase reporter assay, RNA immunoprecipitation, subcellular distribution, qRT-PCR and western blotting. RESULTS: lncRNA KCNQ1OT1 was up-regulated in Rb tissues and Rb cell lines. In addition, the expression of KCNQ1OT1 was negatively correlated with the disease-free survival rate of RB patients. Silencing KCNQ1OT1 could significantly inhibit the RB progression in vivo and in vitro. The analysis of the mechanism of KCNQ1OT1 showed that KCNQ1OT1 can sponge miR-134, and miR-134 may inhibit TRIM44 expression. Moreover, the rescue assays showed that KCNQ1OT1 promoted RB progression by regulating the miR-134/TRIM44 pathway. CONCLUSIONS: The present study indicates that a new KCNQ1OT1/miR-134/TRIM44 pathway regulates Rb progression. It may be used as a potential prognostic marker for Rb.
Assuntos
Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Interferência de RNA , Retinoblastoma/genética , Regiões 3' não Traduzidas , Animais , Biomarcadores Tumorais , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células , Modelos Animais de Doenças , Feminino , Inativação Gênica , Xenoenxertos , Humanos , Hibridização in Situ Fluorescente , Peptídeos e Proteínas de Sinalização Intracelular/genética , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Camundongos , Canais de Potássio de Abertura Dependente da Tensão da Membrana/genética , Retinoblastoma/metabolismo , Retinoblastoma/patologia , Proteínas com Motivo Tripartido/genética , Proteínas com Motivo Tripartido/metabolismoRESUMO
BACKGROUND: Many potential biomarkers have been identified and studied for bladder cancer diagnosis. In this study, we investigated the role of a new biomarker, long noncoding RNA (lncRNA) PCAT6, in bladder cancer diagnosis and prognosis. METHODS AND RESULTS: The lncRNA PCAT6 expression profile of BC is analyzed using the Cancer Genome Atlas Urothelial Bladder Carcinoma (TCGA-BLCA) data. PCAT6 expression level in 106 pairs of BC tissues and adjacent normal tissues was detected and compared using qRT-PCR. Then, the association between PCAT6 expression and clinicopathologic indicators of BC was evaluated. Meanwhile, the prognostic value of PCAT6 was tested using Kaplan-Meier analysis. Additionally, loss-of-function assays were used to explore the effect of PCAT6 on the biological function of BC cells. We identified that the expression level of PCAT6 in BC tissue was higher than that in adjacent normal tissues. And the BC patients have higher serum PCAT6 than that in healthy volunteers. In addition, the expression level of PCAT6 was correlated with tumor size (p = 0.005), differentiation (p = 0.018), TNM stage (p = 0.04), lymph nodes metastasis (p = 0.019), and distant metastasis (p = 0.028). Kaplan-Meier analysis showed that BC patients with high PCAT6 expression had shorter overall survival (OS) and progression-free survival (PFS). The loss-of-function results revealed that the proliferation and viability of BC cells in PCAT6 knockdown groups decreased significantly, compared with the negative control groups. CONCLUSION: Our results demonstrated that PCAT6 might be a potential biomarker for diagnosis and prognosis of BC.
Assuntos
Biomarcadores Tumorais/análise , Carcinoma de Células de Transição/diagnóstico , RNA Longo não Codificante , Neoplasias da Bexiga Urinária/diagnóstico , Adulto , Idoso , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Carcinoma de Células de Transição/patologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , RNA Longo não Codificante/análise , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Neoplasias da Bexiga Urinária/patologiaRESUMO
Piroxicam (PX) is a nonsteroidal anti-inflammatory drug (NSAID) commonly associated with gastrointestinal (GI) injuries, including dyspepsia, heartburn, inflammation, bleeding, ulceration, and life-threatening perforation. The ß-cyclodextrin (ß-CD)-based PX formulation (PX@CD) has been shown to reduce gastric side effects by improving PX's solubility and dissolution rates. However, the solubility of PX can only be increased to a limited extent by ß-CD, due to the low binding constant between PX and ß-CD (â¼100 M-1). As a result, adverse reactions such as epigastric pain and pyrosis are still commonly reported. Cucurbit[7]uril (CB[7]) is a synthetic macrocyclic host compound that binds strongly to various drugs. In this study, we demonstrated that CB[7] forms complexes with PX in the gastric acid environment with a binding constant approximately 70 times higher than that between ß-CD and PX. The PX@CB[7] inclusion complexes exhibited rapid dissolution rates in the gastric environment. In addition, PX@CB[7] showed significantly higher oral bioavailability and maximum concentration (Cmax) compared to PX and PX@CD (1:2.5), resulting in improved anti-inflammatory effects in both mouse and rat models. Moreover, PX@CB[7] (1:2.5) had the least adhesion to the gastric mucosa and caused the mildest gastric side effects in rat models when compared to PX, PX@CD (1:2.5), and PX@CB[7] (1:1). Lastly, CB[7] demonstrated good oral biocompatibility in a subacute toxicity evaluation study. These findings indicate that CB[7] could be used as an excipient to improve treatment effectiveness and decrease adverse reactions in orally administered formulations with a favorable safety profile.
Assuntos
Anti-Inflamatórios não Esteroides , Disponibilidade Biológica , Hidrocarbonetos Aromáticos com Pontes , Imidazóis , Piroxicam , Solubilidade , beta-Ciclodextrinas , Animais , Piroxicam/administração & dosagem , Piroxicam/química , Piroxicam/farmacocinética , Piroxicam/efeitos adversos , Imidazóis/química , Imidazóis/administração & dosagem , Imidazóis/farmacocinética , Imidazóis/efeitos adversos , Hidrocarbonetos Aromáticos com Pontes/química , Hidrocarbonetos Aromáticos com Pontes/administração & dosagem , Hidrocarbonetos Aromáticos com Pontes/farmacocinética , Anti-Inflamatórios não Esteroides/administração & dosagem , Anti-Inflamatórios não Esteroides/química , Anti-Inflamatórios não Esteroides/farmacocinética , Anti-Inflamatórios não Esteroides/efeitos adversos , beta-Ciclodextrinas/química , beta-Ciclodextrinas/administração & dosagem , Masculino , Camundongos , Ratos Sprague-Dawley , Ratos , Liberação Controlada de Fármacos , Administração Oral , Compostos Heterocíclicos com 2 Anéis , Compostos Macrocíclicos , ImidazolidinasRESUMO
Genistein (GEN) is a molecule of great interest as a potent chemopreventive agent against atherosclerosis and cancer. However, the bioavailability of GEN is very low in vivo. Our previous study showed that a GEN derivative, 7-difluoromethyl-5,4'-dimethoxygenistein (dFMGEN) has a better bioavailability than GEN in vivo. In this study, we further evaluated the efficacy of dFMGEN as a candidate for cancer therapy. We demonstrated that dFMGEN treatment decreased the viability of A549 cells in a concentration- and time-dependent manner and induced cell-cycle arrest at the G(1) phase. G(1) phase arrest was correlated with a significant reduction of Cdk4 and cyclin D1 protein level. Further studies showed that cyclin-dependent kinase (Cdk)4 and cyclin D1 protein-level decrease was caused by Cdk inhibitors p15, p21, and p27 level increase, and decreased protein level directly suppressed Rb protein phosphorylation and E2F-1 expression, then cell-cycle progression was arrested. Finally, we also found that dFMGEN has a dosage effect in suppressing tumor growth in vivo, and that dFMGEN was well tolerated by animals. In summary, our results suggest that dFMGEN has therapeutic potential for the treatment of human lung cancer.
Assuntos
Proliferação de Células/efeitos dos fármacos , Pontos de Checagem da Fase G1 do Ciclo Celular/efeitos dos fármacos , Genisteína/análogos & derivados , Neoplasias Pulmonares/tratamento farmacológico , Animais , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Ciclina D1/efeitos dos fármacos , Ciclina D1/metabolismo , Quinase 4 Dependente de Ciclina/efeitos dos fármacos , Quinase 4 Dependente de Ciclina/metabolismo , Relação Dose-Resposta a Droga , Genisteína/administração & dosagem , Genisteína/farmacologia , Genisteína/toxicidade , Humanos , Neoplasias Pulmonares/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Fosforilação/efeitos dos fármacos , Proteína do Retinoblastoma/metabolismo , Fatores de Tempo , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Multidrug resistance is a major obstacle to successful treatment of oral squamous cell carcinoma (OSCC). Lately, we found a novel human gene named tongue cancer chemotherapy resistance-associated protein1 (TCRP1) in the tongue cancer multi-drug resistance cell line (Tca8113/PYM) established by us. In this study, we focus on recombinant expression, purification, and biochemical characterization of TCRP1. After molecular cloning and purification of the gene encoding the 24-kDa protein, a mouse polyclonal antibody against TCRP1 was prepared, and the specialty of the antibody was confirmed by Western blot. The cell proliferation was evaluated by MTS assay and DNA damage was determined by comet assay, the results indicated that this protein especially mediated the cell's resistance to cisplatin; it was associated with its role of providing protection against DNA damage. We also found that TCRP1 expression was increased in cisplatin-resistant carcinoma cell lines (Tca/PYM and A549/DDP), but not in cisplatin-sensitive MDR cell lines (MCF-7/5-Fu), compared with their parental counterparts by Western blot analysis. Immunofluorescence and immunohistochemical analysis showed TCRP1 is mainly expression in cytoplasmic, the Mann-Whitney U test exhibited that TCRP1 positive patients predicted the worst sensitive with cisplatin of OSCC patients. All these findings suggest that TCRP1 is a novel cisplatin-resistant protein which is mainly localized in the cytoplasm and can mediate cisplatin resistance against DNA damage; the expression level of TCRP1 in patients with OSCC may be useful as an indicator of therapeutic efficacy of the sensitivity to cisplatin.
Assuntos
Carcinoma de Células Escamosas/metabolismo , Neoplasias Bucais/metabolismo , Proteínas/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Animais , Antibióticos Antineoplásicos/farmacologia , Especificidade de Anticorpos , Bleomicina/análogos & derivados , Bleomicina/farmacologia , Carcinoma de Células Escamosas/patologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Cisplatino/farmacologia , Dano ao DNA , Resistencia a Medicamentos Antineoplásicos , Fluoruracila/farmacologia , Humanos , Interações Hidrofóbicas e Hidrofílicas , Soros Imunes/química , Camundongos , Camundongos Endogâmicos BALB C , Neoplasias Bucais/patologia , Ligação Proteica , Proteínas/imunologia , Proteínas/isolamento & purificação , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/isolamento & purificaçãoRESUMO
Retinoblastoma (RB) is an intraocular malignancy that mainly affects young children. Previous reports have demonstrated that mutations or the inactivation of the RB1 gene were the main cause of RB; however, disruption of the intracellular signaling pathways following deficiency of RB1 requires further investigation. Based on the Gene Expression Omnibus data and bioinformatics prediction, the present study aimed to investigate the microRNA (miR)3383p/neurooncological ventral antigen 1 (NOVA1) axis in RB. Subsequently, overexpression and knockdown of miR3383p and NOVA1, respectively, were performed to study the role of miR3383p/NOVA1 in the progression of the RB cells. The results demonstrated that overexpression of miR3383p significantly inhibited cell proliferation, migration and invasion, and promoted apoptosis of the RB cells. Moreover, knockdown of NOVA1 showed similar results. A dualluciferase reporter assay and rescue experiments further confirmed the direct binding between miR3383p and NOVA1. Taken together, the results indicated that miR3383p acted as tumor suppressor by targeting the oncogene of NOVA1 in RB, which may serve as potential therapeutic targets in RB.
Assuntos
MicroRNAs/genética , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a Retinoblastoma/genética , Retinoblastoma/genética , Ubiquitina-Proteína Ligases/genética , Adulto , Apoptose/genética , Linhagem Celular Tumoral , Proliferação de Células/genética , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Masculino , Pessoa de Meia-Idade , Mutação/genética , Antígeno Neuro-Oncológico Ventral , Retinoblastoma/patologia , Transdução de Sinais/genéticaRESUMO
OBJECTIVE: We report 3 cases of polycystic ovary syndrome (PCOS) in which the patients had successful pregnancy after repeated implantation failure in at least 8 in vitro fertilization and embryo transfer (IVF-ET) cycles. The patients were treated with gonadotropin-releasing hormone antagonist (GnRH-ant) protocol and gonadotropin-releasing hormone angonist (GnRHa) for triggering ovulation, and successful pregnancy and normal deliveries were achieved after 9 IVT-ET cycles. For young patients with PCOS but a good ovarian reserve and a high ovarian response, treatment with GnRH antagonist protocol and GnRHa alone with appropriate management of the factors that may affect implantation can prevent severe ovarian hyperstimulation syndrome to achieve favorable clinical outcomes.
Assuntos
Implantação do Embrião , Síndrome do Ovário Policístico , Resultado da Gravidez , Transferência Embrionária , Feminino , Fertilização in vitro , Hormônio Liberador de Gonadotropina/agonistas , Hormônio Liberador de Gonadotropina/antagonistas & inibidores , Gonadotropinas , Antagonistas de Hormônios/uso terapêutico , Humanos , Síndrome de Hiperestimulação Ovariana , Ovulação , Indução da Ovulação , GravidezRESUMO
We previously demonstrated that 14-3-3σ was downregulated in 5-fluorouracil (5-Fu)-resistant MCF-7 breast cancer cells (MCF-7/5-Fu). Here, we found that stably enhanced 14-3-3σ expression strengthened the effects of 5-Fu, Mitoxantrone and cDDP. 14-3-3σ stabilised the p53 protein and bound Akt to inhibit its activity and its downstream targets: survivin, Bcl-2 and NF-κB-p50. In addition, decreased p53 expression, but not promoter hypermethylation, was responsible for the downregulation of 14-3-3σ in MCF-7/5-Fu cells. Meanwhile, initial treatments with high concentrations of 5-Fu clearly induced 14-3-3σ and p53 expression in a time-dependent manner. 14-3-3σ-mediated molecular events that synergise with p53 may play important roles in the chemotherapy of breast cancer.