RESUMO
<p><b>OBJECTIVE</b>To investigate the killing effect of herpes simplex virus thymidine kinase gene expression actuated by the promoter of the vascular endothelial growth factor gene on human hepatocellular tumor cells under hypoxic condition.</p><p><b>METHODS</b>Recombinant adenoviral vectors, AdVEGF-tk and AdVEGF-GFP, were constructed with HSV-tk or GFP under the control of VEGF promoter through AdEasy system. Then GFP expression in hepatoma cell line HepG2 and normal liver cell line L02 transfected with AdVEGF-GFP were observed under fluorescence microscope, and the sensitivity to GCV of the AdVEGF-tk-transfected cells under normoxia or hypoxia condition were monitored by MTT method.</p><p><b>RESULTS</b>GFP expression actuated by VEGF promoter was detected in sporadic L02 cells, but in almost all HepG2 cells after transfected with AdVEGF-GFP. With GCV at 10 micro g/ml and MOI at 100, L02 cells were insensitive to GCV under oxic condition, but more than 70% L02 cells were killed under hypoxic condition. Moreover, HepG2 cells infected with AdVEGF-tk showed the increased GCV sensitivity under hypoxia (over 80% killed) as compared with normoxia (over 60% killed) conditions.</p><p><b>CONCLUSION</b>Hypoxia enhances the GCV sensitivity of human hepatocellular tumor cells infected with recombinant AdVEGF-tk under the control of VEGF promoter.</p>
Assuntos
Humanos , Adenoviridae , Genética , Carcinoma Hepatocelular , Patologia , Fatores de Crescimento Endotelial , Genética , Expressão Gênica , Técnicas de Transferência de Genes , Vetores Genéticos , Genética , Hipóxia , Peptídeos e Proteínas de Sinalização Intercelular , Genética , Neoplasias Hepáticas , Patologia , Linfocinas , Genética , Oxigênio , Farmacologia , Regiões Promotoras Genéticas , Fisiologia , Simplexvirus , Timidina Quinase , Genética , Metabolismo , Células Tumorais Cultivadas , Fator A de Crescimento do Endotélio Vascular , Fatores de Crescimento do Endotélio VascularRESUMO
<p><b>OBJECTIVE</b>To investigate the correlation between midkine (MK) protein expression with local infiltration and metastasis in human hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>Immunohistochemical and Western Blot analysis for MK were performed on samples of tumor tissue and the paratumor tissue from HCC and benign liver tumors.</p><p><b>RESULTS</b>The overexpression of MK protein determined by immunohistochemical analysis was similar to that by Western Blot analysis. No specific positivity was detected in either benign liver tumor tissue or normal liver tissue, but most of HCC tissue showed a positive reaction to MK immunostain. No correlation between MK expression and other clinicopathological features in MK negative or positive HCC cases was found. Yet, the overexpression rate of MK protein in HCC with intra-hepatic metastasis was significantly higher than that in HCC without intra-hepatic metastasis.</p><p><b>CONCLUSION</b>In human hepatocellular carcinoma, MK overexpressed at protein level may very well be closely related to local infiltration and metastasis.</p>
Assuntos
Humanos , Western Blotting , Carcinoma Hepatocelular , Metabolismo , Proteínas de Transporte , Metabolismo , Citocinas , Imuno-Histoquímica , Neoplasias Hepáticas , Metabolismo , Patologia , Metástase NeoplásicaRESUMO
0.05). The mean growth values in AdCMV-tk/GCV- and AdVEGF-tk/GCV-treated tumors were significantly lower than those in untreated tumors and AdVEGF-tk/saline-untreated tumors( P