Hepatitis B virus （HBV） biosynthesis is primarily restricted to hepatocytes due to the governing of liver-enriched nuclear receptors （NRs） on viral RNA synthesis. The liver-enriched NR hepatocyte nuclear factor 4...Hepatitis B virus （HBV） biosynthesis is primarily restricted to hepatocytes due to the governing of liver-enriched nuclear receptors （NRs） on viral RNA synthesis. The liver-enriched NR hepatocyte nuclear factor 4α （HNF4α, the key regulator of genes implicated in hepatic glucose metabolism, is also a primary determinant of HBV pregenomic RNA synthesis and HBV replication. Peroxisome proliferator-activated receptor-r coactivator la （PGCla） coactivates and further enhances the effect of HNF4α on HBV biosynthesis. Here, we showed that the acetyltransferase General Control Non-repressed Protein 5 （GCN5） acetylated PGC 1 α, leading to alteration of PGC 1 α from a transcriptionally active state into an inactive state. As a result, the coactivation activity of PGCla on HBV transcription and replication was suppressed. Apparently, an acetylation site mutant of PGC 1 α （PGC 1 αR13） still had coactivation activity as GCN5 could not suppress the coactivation activity of the mutant. Moreover, a catalytically inactive acetyltransferase mutant GCN5m, due to the loss of acetylation activity, failed to inhibit the coactivation function of PGClα in HBV biosynthesis. Our results demonstrate that GCN5, through its acetyltransferase activity, inhibits PGCla-induced enhancement of HBV transcription and replication both in vitro and in vivo.展开更多
Objective:To investigate the effect of Delisheng Injection（得力生注射液 DLS）,a Chinese medicinal compound,DLS combined with cis-platinum（DDP）,an active agent used in lung cancer chemotherapy,on a human highly meta...Objective:To investigate the effect of Delisheng Injection（得力生注射液 DLS）,a Chinese medicinal compound,DLS combined with cis-platinum（DDP）,an active agent used in lung cancer chemotherapy,on a human highly metastatic giant lung carcinoma cell line PGCL3.Methods:The suspended PGCL3 cells at10;/mL cultured in 96-well tissue culture plates were divided into 4 groups:DLS treatment group（2 μL/mL,5 μL/mL,10 μL/mL,25 μL/mL）,DDP treatment group（1 μg/mL,2 μg/mL,5 μg/mL,15 μg/mL）,combined DLS with DDP treatment group（DLS:DDP 2 μL/mL:1 μg/mL,5 μL/mL:2 μg/mL,10 μL/mL:5 μg/mL,25 μL/mL:15 μg/mL）and a control group.The cytotoxicity of DLS with different concentrations（2 μL/mL,5 μL/mL,10 μL/mL,25 μL/mL）on PGCL3 cells was determined by 3-（4,5）-dimethylthiahiazo（-z-y1）-3,5-di-phenytetrazoliumromide（MTT） assay.Effect of DLS on adhesion of PGCL-3 cells was tested by cell-matrigel adhesion assay.Chemotactic movement model of transwell camerula was used to determine the effect of DLS on invasion and migration of PGCL-3 cells.Results:Compared with the control group,DLS（2 μL/mL,5 μL/mL,10 μL/mL,25 μL/mL） could significantly decrease cell proliferation,adhesion,invasion and migration abilities（P<0.05）.Cell adhesion,invasion and migration abilities were significantly decreased after combination treatment of DLS:DDP（2 μL/mL:1 μg/mL,5 μL/mL:2 μg/mL,10 μL/mL:5 μg/mL,25 μL/mL:15 μg/mL） compared with DDP single-agent treatment（1 μg/mL,2 μg/mL,5 μg/mL,15 μg/mL,P<0.05）,respectively.Conclusions:DLS single-agent has a satisfying inhibition effect in PGCL3 cell line and DLS might enhance the inhibition effect of DDP on cancer metastasis.Our research provided a experimental basis about the treatment on highly metastatic lung caner.展开更多
基金supported by grants from the National Major Science and Technology Special Projects for Infectious Diseases of China (2012ZX10004503-008, 2012ZX10001006-002,and 2012ZX10002006-002)
文摘Hepatitis B virus （HBV） biosynthesis is primarily restricted to hepatocytes due to the governing of liver-enriched nuclear receptors （NRs） on viral RNA synthesis. The liver-enriched NR hepatocyte nuclear factor 4α （HNF4α, the key regulator of genes implicated in hepatic glucose metabolism, is also a primary determinant of HBV pregenomic RNA synthesis and HBV replication. Peroxisome proliferator-activated receptor-r coactivator la （PGCla） coactivates and further enhances the effect of HNF4α on HBV biosynthesis. Here, we showed that the acetyltransferase General Control Non-repressed Protein 5 （GCN5） acetylated PGC 1 α, leading to alteration of PGC 1 α from a transcriptionally active state into an inactive state. As a result, the coactivation activity of PGCla on HBV transcription and replication was suppressed. Apparently, an acetylation site mutant of PGC 1 α （PGC 1 αR13） still had coactivation activity as GCN5 could not suppress the coactivation activity of the mutant. Moreover, a catalytically inactive acetyltransferase mutant GCN5m, due to the loss of acetylation activity, failed to inhibit the coactivation function of PGClα in HBV biosynthesis. Our results demonstrate that GCN5, through its acetyltransferase activity, inhibits PGCla-induced enhancement of HBV transcription and replication both in vitro and in vivo.
基金Supported by the Project of Science and Technology Development in Shaanxi Province of China(No.2008KG10-01)
文摘Objective:To investigate the effect of Delisheng Injection（得力生注射液 DLS）,a Chinese medicinal compound,DLS combined with cis-platinum（DDP）,an active agent used in lung cancer chemotherapy,on a human highly metastatic giant lung carcinoma cell line PGCL3.Methods:The suspended PGCL3 cells at10;/mL cultured in 96-well tissue culture plates were divided into 4 groups:DLS treatment group（2 μL/mL,5 μL/mL,10 μL/mL,25 μL/mL）,DDP treatment group（1 μg/mL,2 μg/mL,5 μg/mL,15 μg/mL）,combined DLS with DDP treatment group（DLS:DDP 2 μL/mL:1 μg/mL,5 μL/mL:2 μg/mL,10 μL/mL:5 μg/mL,25 μL/mL:15 μg/mL）and a control group.The cytotoxicity of DLS with different concentrations（2 μL/mL,5 μL/mL,10 μL/mL,25 μL/mL）on PGCL3 cells was determined by 3-（4,5）-dimethylthiahiazo（-z-y1）-3,5-di-phenytetrazoliumromide（MTT） assay.Effect of DLS on adhesion of PGCL-3 cells was tested by cell-matrigel adhesion assay.Chemotactic movement model of transwell camerula was used to determine the effect of DLS on invasion and migration of PGCL-3 cells.Results:Compared with the control group,DLS（2 μL/mL,5 μL/mL,10 μL/mL,25 μL/mL） could significantly decrease cell proliferation,adhesion,invasion and migration abilities（P<0.05）.Cell adhesion,invasion and migration abilities were significantly decreased after combination treatment of DLS:DDP（2 μL/mL:1 μg/mL,5 μL/mL:2 μg/mL,10 μL/mL:5 μg/mL,25 μL/mL:15 μg/mL） compared with DDP single-agent treatment（1 μg/mL,2 μg/mL,5 μg/mL,15 μg/mL,P<0.05）,respectively.Conclusions:DLS single-agent has a satisfying inhibition effect in PGCL3 cell line and DLS might enhance the inhibition effect of DDP on cancer metastasis.Our research provided a experimental basis about the treatment on highly metastatic lung caner.
