RESUMEN
Lung cancer mortality remains high, and only approximately 15% of patients with non-small cell lung cancer survive for more than five years. The purpose of this research was to investigate the prognostic value and biological functions of G protein regulated inducer of neuritis outgrowth 1(GPRIN1) in lung cancer. We used the Kaplan-Meier method to analyze the correlation between GPRIN1 and overall survival, and performed Cox regression to determine whether GPRIN1 might be an independent predictive factor for lung adenocarcinoma prognosis. qRT-PCR and Western blot assays were conducted to detect GPRIN1 expression in lung cancer cells and normal control cells. To detect the functional effects of knockdown/overexpression of GPRIN1 on lung cancer cells, we performed CCK-8, colony formation and Transwell assays. Through the Kaplan-Meier method, we found that GPRIN1 expression correlated with overall survival and adverse prognosis, and Cox regression indicated that GPRIN1 is as an independent predictive factor for lung adenocarcinoma. Furthermore, the mRNA and protein expression levels of GPRIN1 in lung cancer cells were markedly higher than those in normal cells. Downregulation of GPRIN1significantly decreased cell viability, colony formation, the number of invasive and migrating cells, and levels of epithelial-mesenchymal transition-related proteins in A549 cells. Overexpression of GPRIN1showed the opposite effect in Calu-1 cells. Together, these results indicated that GPRIN1 facilitates lung cancer proliferation and migration, possibly by affecting the epithelial-mesenchymal transition of lung cancer cells, suggesting that GPRIN1may be used as an effective target for the treatment of lung cancer.
Asunto(s)
Neoplasias Pulmonares , Carcinoma de Pulmón de Células no Pequeñas/genética , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Transición Epitelial-Mesenquimal , Regulación Neoplásica de la Expresión Génica , Humanos , Neoplasias Pulmonares/genética , Invasividad Neoplásica/genética , Proteínas del Tejido Nervioso , Proyección Neuronal , Receptores de N-Metil-D-AspartatoRESUMEN
Tenascin-C (TN-C) is a key component of extracellular matrix (ECM) and its expression process is poorly understood during rheumatic heart valvular disease (RHVD). In this study, we found that interferon (IFN)-gamma, tumour necrosis factor (TNF)-alpha and TN-C concentrations in patients with RHVD were significantly higher than in normal controls. More IFN-gamma receptors and TNF receptors were found being expressed on rheumatic aortic valves interstitial cells than on non-rheumatic ones and their expression was patients' sera dependent. Antibodies neutralizing IFN-gamma or TNF-alpha could attenuate patients' sera-induced TN-C transcription by isolated rheumatic aortic valves interstitial cells. By application with different protein kinase inhibitors, we found that combined with cyclic strain, TNF-alpha and IFN-gamma induced TN-C transcription through the RhoA/ROCK signalling pathway. At the same time, p38 mitogen-activated protein kinase was involved in TNF-alpha and IFN-gamma induced TN-C transcription. TNF-alpha also increased TN-C mRNA level by additional PKC and ERK 1/2 activation. Our finding revealed a new insight into ECM remodelling during RHVD pathogenesis and new mechanisms involved in the clinical anti-IFN-gamma and anti-TNF-alpha therapy.
Asunto(s)
Válvula Aórtica/metabolismo , Citocinas/fisiología , Cardiopatía Reumática/metabolismo , Tenascina/biosíntesis , Adulto , Válvula Aórtica/anomalías , Válvula Aórtica/patología , Células Cultivadas , Femenino , Corazón Auxiliar , Humanos , Interferón gamma/sangre , Masculino , Persona de Mediana Edad , ARN Mensajero/genética , Receptores de Interferón/metabolismo , Receptores del Factor de Necrosis Tumoral/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Cardiopatía Reumática/sangre , Cardiopatía Reumática/patología , Estrés Mecánico , Tenascina/sangre , Tenascina/genética , Activación Transcripcional/inmunología , Factor de Necrosis Tumoral alfa/sangre , Adulto Joven , Receptor de Interferón gammaRESUMEN
A simply hemoglobin (Hb) molecularly imprinted polymer (MIP) was prepared using Hb as the imprinted molecule, acrylamide as the functional monomer and cross-linked chitosan beads as the supporting matrix. The MIP was achieved by entrapment of the selective soft polyacrylamide gel in the pores of the cross-linked chitosan beads by letting acrylamide monomer and the protein diffuse into the pores of chitosan beads before starting the polymerization. The chitosan beads were freed from the surrounding polyacrylamide gel by washing. The Langmuir and Freundlich adsorption models were applied to describe the equilibrium isotherms. Langmuir analysis showed that an equal class of adsorption was formed in the MIP and the adsorption equilibrium constant and the maximum adsorption capacity were evaluated. The MIP has much higher adsorption capacity for Hb than the non-imprinted polymer with the same chemical composition, and the MIP also has a higher selectivity for the imprinted molecule. The MIP can be reused in an easy way and the reproduction coefficient was approximately 100% at low concentration.
