Effects of hepatitis C virus genotype 1b core protein on the gene expressions of Bax and Bcl-2 in HepG2 cells
Lei Chuang
Abstract
Lei Chuang
Abstract
Objective To investigate the effects of hepatitis C virus genotype 1b core protein on the expression of Bax and Bcl-2 in the HepG2 cell and to study the relationship of core protein and cell apoptosis.Methods The gene of HCV-1b-C was amplified by RT-PCR,and then it was inserted into pcDNA3.1(-) to construct the expression plasmid pcDNA3.1(-)/HCV-1b-C by double enzyme-cut.To transfect the HepG2 cell with recombination expression vector,pcDNA3.1(-)/ HCV-1b-C,by lipofectamine.The expression of HCV C mRNA and protein was detected by RT-PCR and Western Blot respectively.And then,detect the effects of HCV-1b-C on the expression of Bax and Bcl-2 gene in the HepG2 cell by RT-PCR and Western Blot,and set groups transfect with the plasmid pcDNA3.1(-) and normal groups as control.Results The expression vector pcDNA3.1(-)/ HCV-1b-C was constructed successfully;And then,HCV C protein was expressed in the HepG2 cell line by transfection;For the corresponding mRNA and protein expression of Bax were lower in the groups transfected with HCV-1b-C compare to those in the groups transfected with the empty plasmid and normal groups(P0.01);For the corresponding mRNA and protein expression of Bcl-2 were higher in the groups transfected with HCV-1b-C than those of in groups transfected with the empty plasmid and normal groups(P0.01).Conclusion The expression of genotype 1b HCV C in the HepG2 cell can reduce the expression of Bax and increase the expression of Bcl-2,leading to decrease in the ratio of Bax /Bcl-2,considering one of the mechanism of HepG2 cell escape from cell apoptosis.
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Objective To investigate the effects of hepatitis C virus genotype 1b core protein on the expression of Bax and Bcl-2 in the HepG2 cell and to study the relationship of core protein and cell apoptosis.Methods The gene of HCV-1b-C was amplified by RT-PCR,and then it was inserted into pcDNA3.1(-) to construct the expression plasmid pcDNA3.1(-)/HCV-1b-C by double enzyme-cut.To transfect the HepG2 cell with recombination expression vector,pcDNA3.1(-)/ HCV-1b-C,by lipofectamine.The expression of HCV C mRNA and protein was detected by RT-PCR and Western Blot respectively.And then,detect the effects of HCV-1b-C on the expression of Bax and Bcl-2 gene in the HepG2 cell by RT-PCR and Western Blot,and set groups transfect with the plasmid pcDNA3.1(-) and normal groups as control.Results The expression vector pcDNA3.1(-)/ HCV-1b-C was constructed successfully;And then,HCV C protein was expressed in the HepG2 cell line by transfection;For the corresponding mRNA and protein expression of Bax were lower in the groups transfected with HCV-1b-C compare to those in the groups transfected with the empty plasmid and normal groups(P0.01);For the corresponding mRNA and protein expression of Bcl-2 were higher in the groups transfected with HCV-1b-C than those of in groups transfected with the empty plasmid and normal groups(P0.01).Conclusion The expression of genotype 1b HCV C in the HepG2 cell can reduce the expression of Bax and increase the expression of Bcl-2,leading to decrease in the ratio of Bax /Bcl-2,considering one of the mechanism of HepG2 cell escape from cell apoptosis.
Key concepts: Transfection, Lipofectamine, Molecular biology, Plasmid, Expression vector, Hepatitis C virus, Western blot, Gene