ABSTRACT
FXYD6, FXYD domain containing ion transport regulator 6, has been reported to affect the activity of Na(+)/K(+)-ATPase and be associated with mental diseases. Here, we demonstrate that FXYD6 is up-regulated in hepatocellular carcinoma (HCC) and enhances the migration and proliferation of HCC cells. Up-regulation of FXYD6 not only positively correlates with the increase of Na(+)/K(+)-ATPase but also coordinates with the activation of its downstream Src-ERK signaling pathway. More importantly, blocking FXYD6 by its functional antibody significantly inhibits the growth potential of the xenografted HCC tumors in mice, indicating that FXYD6 represents a potential therapeutic target toward HCC. Altogether, our results establish a critical role of FXYD6 in HCC progression and suggest that the therapy targeting FXYD6 can benefit the clinical treatment toward HCC patients.
Subject(s)
Animals , Female , Humans , Antibodies, Monoclonal , Pharmacology , Antineoplastic Agents , Pharmacology , Carcinoma, Hepatocellular , Drug Therapy , Metabolism , Cell Line, Tumor , Cell Movement , Cell Proliferation , HEK293 Cells , Ion Channels , Metabolism , Liver Neoplasms , Drug Therapy , Metabolism , Mice, Inbred BALB C , Mice, Nude , Sodium-Potassium-Exchanging ATPase , Metabolism , Tumor Burden , Xenograft Model Antitumor AssaysABSTRACT
CD146 is a newly identified endothelial biomarker that has been implicated in angiogenesis. Though in vitro angiogenic function of CD146 has been extensively reported, in vivo evidence is still lacking. To address this issue, we generated endothelial-specific CD146 knockout (CD146(EC-KO)) mice using the Tg(Tek-cre) system. Surprisingly, these mice did not exhibit any apparent morphological defects in the development of normal retinal vasculature. To evaluate the role of CD146 in pathological angiogenesis, a xenograft tumor model was used. We found that both tumor volume and vascular density were significantly lower in CD146(EC-KO) mice when compared to WT littermates. Additionally, the ability for sprouting, migration and tube formation in response to VEGF treatment was impaired in endothelial cells (ECs) of CD146(EC-KO) mice. Mechanistic studies further confirmed that VEGF-induced VEGFR-2 phosphorylation and AKT/p38 MAPKs/NF-κB activation were inhibited in these CD146-null ECs, which might present the underlying cause for the observed inhibition of tumor angiogenesis in CD146(EC-KO) mice. These results suggest that CD146 plays a redundant role in physiological angiogenic processes, but becomes essential during pathological angiogenesis as observed in tumorigenesis.
Subject(s)
Animals , Female , Male , Mice , CD146 Antigen , Genetics , Metabolism , Cells, Cultured , Endothelial Cells , Cell Biology , Metabolism , Fibrosarcoma , Metabolism , Pathology , Melanoma, Experimental , Metabolism , Pathology , Mice, Inbred C57BL , Mice, Knockout , NF-kappa B , Metabolism , Neovascularization, Physiologic , Phosphorylation , Proto-Oncogene Proteins c-akt , Metabolism , Retinal Vein , Pathology , Signal Transduction , Transplantation, Homologous , Vascular Endothelial Growth Factor A , Pharmacology , Vascular Endothelial Growth Factor Receptor-2 , MetabolismABSTRACT
During lymphocyte development antigen receptor genes undergo V(D)J recombination to obtain antigen binding specificity and diversity. This process is not only controlled by genetic factors, such as tissue- and stage- specificity of RAG-1/2 protein, germline transcriptional activity and ACEs, but also regulated at epigenetic level. The chromatin accessibility of recombinase is associated with the chromatin configuration around the targeted gene segments. Thus, activation of V(D)J recombination requires the recruitment of remodeling complexes for changing the accessibility in the localized chromatin. Moreover, docking of remodeling complexes, which serve for creating active chromatin environment, relies on certain patterns of chromatin modification. Some recent findings regarding epigenetic regulation mechanisms in V(D)J recombination, such as CpG methylation, histone modification, nucleosome remodeling and nuclear topology were reviewed.
ABSTRACT
A mAb T2-2 was generated using hybridoma techniques, and its target was identified as a 46 ku-cytokeratin (CK), based on biochemical study and a completely overlapped binding pattern of mAb T2-2 with anti-pan-CKs antibodies. An epithelia-specificity of the mAb T2-2 was determined by screening 68 human normal and 65 tumor tissues using immunohistochemistry. Unlike most of anti-CKs antibodies, the mAb T2-2 recognized a mono-specific epitope only expressed on the 46 ku CK, suggesting that mAb T2-2 is superior to most anti-CKs antibodies that cross-reacted with many different kinds of CKs. In addition, it was found that the mAb T2-2 was multipurpose with a broad applicability to ELISA, immunohistochemistry, immunofluorescence, Western blotting, and was also compatible with various fixation reagents. These results strongly indicate that the mAb T2-2 has potential applications for studying CKs function and for diagnosis of tumor and other disorders.
ABSTRACT
To express human single chain antibody to collagenase IV as soluble proteins secreted by pichia pastoris, a recombinant vector scFv-pPIC9 was constructed, and then transformed into pichia pastoris GS115 by electuoporation, The transformants were selected by DNA hybridization and cultured in the media with methanlo. Soluble scFv were secreted into culturesupernatant and characterized by SDS-PAGE and Western Blot. Our results showed that the secreting of soluble scFv in pichia pastoris was as high as 20mg/L. The soluble scFv has similar immuno-activity to its counterpart produced in bacteria, and it is more easily and efficiently purified.
ABSTRACT
To express human single chain antibody to collagenase IV as soluble proteins secreted by pichia pastoris, a recombinant vector scFv pPIC9 was constructed, and then transformed into pichia pastoris GS115 by electuoporation, The transformants were selected by DNA hybridization and cultured in the media with methanlo. Soluble scFv were secreted into culturesupernatant and characterized by SDS PAGE and Western Blot. Our results showed that the secreting of soluble scFv in pichia pastoris was as high as 20mg/L. The soluble scFv has similar immuno activity to its counterpart produced in bacteria, and it is more easily and efficiently purified.
ABSTRACT
Objective To analyze 5 patients suptered heart failure and ventricular tachycardia((VT).)Methods The Implantable Biventricular pacing cardioverter defibrillator was used to analyzed 5 patients(4 with primary dilated cardiomyopathy and 1 with coronary heart disease) suffered from heart failure and ventricular tachycardia(VT).Results 2 cases had history of syncope,4 patients were implanted INSYNC 7272 implantable cardioverter defibrillator(ICD) and 1 patient was implanted V-350 ICD.During the 1~14 month follow-up period,all the cardio functions were improved without VT or syncope.1 patient with VT and VF was detected and terminated by ICD.The patient was saved.Conclusions Implantable biventricular pacing cardioverter defibrillator is an effective approach to treat sudden cardiac death,mortal ventricular arrhythmia and heart failure.