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1.
Journal of Korean Medical Science ; : 1215-1223, 2016.
Artículo en Inglés | WPRIM | ID: wpr-69708

RESUMEN

Orthodenticlehomeobox 1 (OTX1) overexpression had previously been associated with the progression of several tumors. The present study aimed to determine the expression and role of OTX1 in human hepatocellular carcinoma (HCC). The expression level of OTX1 was examined by quantitative real-time PCR (qRT-PCR) in 10 samples of HCC and paired adjacent non-cancerous tissues, and by immunohistochemistry (IHC) analysis in 128 HCC samples and matched controls. The relationship between OTX1 expression and the clinicopathological features werealso analyzed. Furthermore, the effects of OTX1 knockdown on cell proliferation and migration were determined in HCC cell lines. Axenograft mouse model was also established to investigate the role of OTX1 in HCC tumor growth. TheqRT-PCR and IHC analyses revealed that OTX1 was significantly elevated in HCC tissues compared with the paired non-cancerous controls. Expression of OTX1 was positively correlated with nodal metastasis status (P = 0.009) and TNM staging (P = 0.001) in HCC tissues. In addition, knockdown of OTX1 by shRNA significantly inhibited the proliferation and migration, and induced cell cycle arrest in S phase in vitro. Tumor growth was markedly inhibited by OTX1 silencing in the xenograft. Moreover, OTX1 silencing was causable for the decreased phosphorylation level of ERK/MAPK signaling. In conclusion, OTX1 contributes to HCC progression possibly by regulation of ERK/MAPK pathway. OTX1 may be a novel target for molecular therapy towards HCC.


Asunto(s)
Anciano , Animales , Femenino , Humanos , Masculino , Ratones , Persona de Mediana Edad , Western Blotting , Carcinoma Hepatocelular/metabolismo , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Progresión de la Enfermedad , Regulación Neoplásica de la Expresión Génica , Inmunohistoquímica , Hígado/metabolismo , Neoplasias Hepáticas/metabolismo , Metástasis Linfática , Sistema de Señalización de MAP Quinasas , Ratones Endogámicos BALB C , Ratones Desnudos , Estadificación de Neoplasias , Factores de Transcripción Otx/antagonistas & inhibidores , Fosforilación , Interferencia de ARN , Reacción en Cadena en Tiempo Real de la Polimerasa , Puntos de Control de la Fase S del Ciclo Celular , Trasplante Heterólogo
2.
Biomedical and Environmental Sciences ; (12): 215-219, 2014.
Artículo en Inglés | WPRIM | ID: wpr-270611

RESUMEN

In this study, a new parameter, S phase cell percentage (S fraction) normalized BrdU (SFN-BrdU) incorporation rate, was introduced to detect S arrest. The results showed a positive linear correlation between the BrdU incorporation rate and the S fraction in unperturbed 16HBE cells. Theoretical analysis indicated that only S arrest could result in a decrease in the SFN-BrdU incorporation rate. Additionally, the decrease in SFN-BrdU incorporation rate and the activation of DNA damage checkpoints further demonstrated that S arrest was induced by diethyl sulfate treatment of 16HBE cells. In conclusion, SFN-BrdU incorporation rate can be used to detecting S arrest.


Asunto(s)
Humanos , Bromodesoxiuridina , Farmacocinética , Proliferación Celular , Daño del ADN , Células Epiteliales , Biología Celular , Fase S , Puntos de Control de la Fase S del Ciclo Celular
3.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 497-503, 2014.
Artículo en Inglés | WPRIM | ID: wpr-351050

RESUMEN

Icaritin, a prenylflavonoid derivative from Epimedium Genus, has been shown to exhibit many pharmacological and biological activities. However, the function and the underlying mechanisms of icaritin in human non-small cell lung cancer have not been fully elucidated. The purpose of this study was to investigate the anticancer effects of icaritin on A549 cells and explore the underlying molecular mechanism. The cell viability after icaritin treatment was tested by MTT assay. The cell cycle distribution, apoptosis and reactive oxygen species (ROS) levels were analyzed by flow cytometry. The mRNA and protein expression levels of the genes involved in proliferation and apoptosis were respectively detected by RT-PCR and Western blotting. The results demonstrated that icaritin induced cell cycle arrest at S phase, and down-regulated the expression levels of S regulatory proteins such as Cyclin A and CDK2. Icaritin also induced cell apoptosis characterized by positive Hoechst 33258 staining, accumulation of the Annexin V-positive cells, increased ROS level and alteration in Bcl-2 family proteins expression. Moreover, icaritin induced sustained phosphorylation of ERK and p38 MAPK. These findings suggested that icaritin might be a new potent inhibitor by inducing S phase arrest and apoptosis in human lung carcinoma A549 cells.


Asunto(s)
Humanos , Antineoplásicos Fitogénicos , Farmacología , Apoptosis , Línea Celular Tumoral , Flavonoides , Farmacología , Neoplasias Pulmonares , Quimioterapia , Metabolismo , Patología , Sistema de Señalización de MAP Quinasas , Proteínas de Neoplasias , Especies Reactivas de Oxígeno , Metabolismo , Puntos de Control de la Fase S del Ciclo Celular
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