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The effect of LIGHT and IFN-γ gene in the apoptosis of HepG₂cells

Other Titles
 HepG2 세포의 세포사멸에 미치는 LIGHT 유전자와 IFN-γ유전자의 영향 
Authors
 오력군 
Issue Date
2009
Description
Dept. of medicine/박사
Abstract
[한글]
[영문] 【Objective】To study the better transfection method in the HepG2 cells mediated by Cationic liposome with LIGHT gene and combined with LIGHT and IFN-γ genes, and the effect of LIGHT and IFN-γ gene in the apoptosis of HepG2 cells.【Methods】To clone full-length ORF of LIGHT and IFN-γ gene. Dividing HepG2 cells into three groups: the control, solo transfection of LIGHT gene and combined transfection of LIGHT and IFN-γ genes, then transfect HepG2 cells mediated by Cationic liposome with pcDNA4C-LIGHT cDNA and pcDNA4His?MaxC-hIFN-γ cDNA extracted from E.coli JM-109, to cellect HepG2 cells on 12 hours,24 hours and 48 hours after transfection respectively .To investigate the apoptosis of HepG2 cells and the expression of cell factors, Fas 、FasL、Survivin、Bcl-2、Caspase-3 and Caspase-8 with flow cytometry. 【Results】After transfection, the apoptosis rate of HepG2 cells was increased with the prolonged time , and the apoptosis rate of the solo transfection group was higher than the control group, while combined group were higher than the solo transfections group(P<0.01). The expression of Fas and FasL were increased in HepG2 cells , but the expression of FasL was lower than that of Fas. The expression of Fas in solo transfection group is higher than that in control group, and the combined group is highest(P<0.01).The expression of FasL in the combined group was higher than that in the solo transfection group on 12 hours and 24 huors, but was lower than that in the solo transfection group on 48 hours after transfection(P<0.01).The apoptosis of HepG2 cells increased as time prolonged ,and the apoptosis of combined group was higher than that of the solo transfection group(P<0.01).The expression of Caspase-3 was gradually increased in the control group、solo group and combined group. On the contratory, the expression of Survivin of decreased gradually. After transfection, the expression of Bcl-2 and Caspase-8 were obviously different than the control group. 【Conclusion】LIGHT and INF-γ genes successfully transfected HepG2 cells mediated by Cationic liposome, pcDNA4/HisMaxC is a stable vector for HepG2 cell apoptosis study. The pcDNA4/HisMax-EGFP identify tansfection efficiency is an ideal method. The possible pathway of LIGHT gene induced HepG2 cell apoptosis is death receptor pathway. LIGHT and INF-γ have synergistic action in HepG2 apoptosis through the down regulation of Bcl-2 expression.
Files in This Item:
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Appears in Collections:
7. Others (기타) > Others (기타) > 5. Others
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/137360
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