RESUMO
Angiopoietin-like protein (ANGPTL) 4 is a key factor in the regulation of lipid and glucose metabolism in metabolic diseases. ANGPTL4 is highly expressed in various cancers, but the regulation of energy metabolism in tumours remains to be determined. This study explored the role of ANGPTL4 in aerobic glycolysis, glutamine consumption and fatty acid oxidation in nonsmall cell lung cancer (NSCLC) cells. Two NSCLC cell lines (A549 and H1299) were used to investigate the role of ANGPTL4 in energy metabolism by tracer techniques and with Seahorse XF technology in ANGPTLs4 knockdown cells. RNA microarrays and specific inhibitors were used to identify targets in ANGPTLs4-overexpressing cells. The results showed that knockdown of ANGPTLs4 could inhibit energy metabolism and proliferation in NSCLC. ANGPTLs4 had no significant effect on glycolysis but affected glutamine consumption and fatty acid oxidation. Knockdown of ANGPTLs4 also significantly inhibited tumour metastasis and energy metabolism in mice and had a weak effect on glycolysis. RNA microarray analysis showed that ANGPTLs4 significantly affected glutaminase (GLS) and carnitine palmitoyl transferase 1 (CPT1). ANGPTLs4-overexpressing cells were exposed to a glutamine deprivation environment, and cell proliferation and energy metabolism were significantly decreased but still differed from normal NSCLC cells. Treatment of ANGPTLs4-overexpressing cells with GLS and CPT1 inhibitors simultaneously prevented the regulatory effects on cell proliferation and energy metabolism. ANGPTLs4 could promote glutamine consumption and fatty acid oxidation but not glycolysis or accelerate energy metabolism in NSCLC.
Assuntos
Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Animais , Carcinoma Pulmonar de Células não Pequenas/patologia , Linhagem Celular Tumoral , Proliferação de Células/genética , Ácidos Graxos/metabolismo , Glutamina/metabolismo , Glicólise , Neoplasias Pulmonares/patologia , CamundongosRESUMO
To clone mouse phage antibodies against H-Y antigen from a phage antibody library, three cycles of affinity enrichment of the mouse phage antibody library with male spleen cells and two cycles of nonspecific absorption with female spleen cells were performed. The presence of mouse Fab on the phage surface was determined by ELISA and sequence analysis. 9 of 15 strains can bind to male spleen cells with the specific activity. Recombination rate of the phage antibody library clones is 60%. Sequence analysis of the PCR products of plasmid DNA of E5 clones show VH and Vkappa had common characteristics shared by other known variable region of antibodies. The mouse phage Fab antibody could be used for identifying H-Y antigen, and for the development of sex determination of early embryos in mammals.
Assuntos
Animais , Feminino , Masculino , Camundongos , Sequência de Bases , Clonagem Molecular , Ensaio de Imunoadsorção Enzimática , Antígeno H-Y , Fragmentos Fab das Imunoglobulinas , Genética , Alergia e Imunologia , Isoanticorpos , Genética , Alergia e Imunologia , Dados de Sequência Molecular , Biblioteca de PeptídeosRESUMO
Objective To investigate the effect of low frequency electric deep brain stimulation on amygdale kindling in rats.Methods The amygdale kinkling model of rats was established by operation on the brain.The effects of low frequency deep brain electric stimulation used alone or in combination with anti-epilepsy drugs were ob- served in terms of severity of seizure attack reflected by Racine's scale and afterdischarge duration recorded in electro- encephalogram.Results Fifteen minutes of low frequency electric stimulation at 1 Hz and 100 to 350?A effective- ly inhibited amygdale kindling as demonstrated by a significant decrease of afterdischarge duration,and decreased the severity of seizure attack (P
RESUMO
Objective To invesgate the effect of P-glycoprotein(PGP)inhibitor,verapamil,on electrobiological activity and seizure behavior in phenytoin-carbamazepine(PHT-CBZ)resistant rats.Methods The model of medically intractable epilepsy was established by kindling of amygdale. Verapamil was applied to PHT-CBZ resistant rats,followed by the observation on after discharge threshold (ADT),after discharge duration(ADD)and seizure activity.Results Compared with the control group, the ADT was higher in PHT-CBZ resistant rats peritoneally injected with verapamil((238.0?32.2)?A vs (177.0?23.3)?A,P
