CAJ | 학술논문

目的观察ARHI基因转染PANC1细胞后对细胞凋亡和自噬相关基因mRNA表达谱的影响.方法采用脂质体法将表达ARHI基因的质粒pIRES2-EGFP-ARHI、空质粒pIRES2-EGFP转染胰腺癌PANC1细胞.采用基因芯片RT2ProfilerTM PCR Array行实时定量PCR,分析转染细胞的基因表达谱,包括84个与凋亡和自噬相关基因.结果 ARHI基因转染组PANC1细胞有9个基因mRNA表达下调,38个基因mRNA表达上调,37个基因mRNA表达变化无意义.在与凋亡相关的基因中有8个促凋亡基因表达显著上调(＞6倍),主要为TNFR/TRFSR家族基因(TNFSF8、TNFRSF10B、TNFRSF11B、TNFRSF9)、CIDE家族基因(DFFA)、CASP家族基因(CASP10、CASP8)和死亡结构域家族基因(DAPK1),其中以DAPK1上调尤为明显,达42.83倍;抗凋亡基因中3个基因(CD27、BCL2L10、BIRC4)表达显著上调(＞6倍),3个基因(BCL2、BAD、BAG4)表达轻度上调(＞2倍),1个基因(BCL2L1)表达轻度下调(＜-2倍).在与自噬相关的基因中3个促自噬基因(TNFRSF10B、DAPK1、CASP10)表达显著上调(＞6倍),4个基因(TNFRSF10A、FADD、TP53、TP53 BP2)表达轻度上调(＞2倍);3个抑制自噬基因(BCL2、CASP8、FAS)表达轻度上调(＞2倍),1个基因(MCL1)表达轻度下调(＜-2倍).结论 ARHI基因显著上调细胞凋亡及自噬重要调控基因Caspase-8和DAPK1.
목적 관찰ARHI기인전염PANC1세포후대세포조망화자서상관기인mRNA표체보적영향.방법 채용지질체법장표체ARHI기인적질립pIRES2-EGFP-ARHI、공질립pIRES2-EGFP전염이선암PANC1세포.채용기인심편RT2ProfilerTM PCR Array행실시정량PCR,분석전염세포적기인표체보,포괄84개여조망화자서상관기인.결과 ARHI기인전염조PANC1세포유9개기인mRNA표체하조,38개기인mRNA표체상조,37개기인mRNA표체변화무의의.재여조망상관적기인중유8개촉조망기인표체현저상조(＞6배),주요위TNFR/TRFSR가족기인(TNFSF8、TNFRSF10B、TNFRSF11B、TNFRSF9)、CIDE가족기인(DFFA)、CASP가족기인(CASP10、CASP8)화사망결구역가족기인(DAPK1),기중이DAPK1상조우위명현,체42.83배;항조망기인중3개기인(CD27、BCL2L10、BIRC4)표체현저상조(＞6배),3개기인(BCL2、BAD、BAG4)표체경도상조(＞2배),1개기인(BCL2L1)표체경도하조(＜-2배).재여자서상관적기인중3개촉자서기인(TNFRSF10B、DAPK1、CASP10)표체현저상조(＞6배),4개기인(TNFRSF10A、FADD、TP53、TP53 BP2)표체경도상조(＞2배);3개억제자서기인(BCL2、CASP8、FAS)표체경도상조(＞2배),1개기인(MCL1)표체경도하조(＜-2배).결론 ARHI기인현저상조세포조망급자서중요조공기인Caspase-8화DAPK1.
Objective To investigate the effect of ARHI transfection on the apoptosis and autophagy related gene expression profile of PANC1 cells.Methods Plasmids pIRES2-EGFP-ARHI which expressing ARHI and empty plasmid pIRES2-EGFP were transfected into PANC1 cells.Expression profile,including 84 apoptosis and autophagy related genes was detected by using quantitative real-time PCR based RT2Profiler TM PCR Array.Results In PANC1 cells transfected with pIRES2-EGFP-ARHI,the expression of mRNA of 9 genes were down-regulated,and 38 were up-regulated,while 37 were not significantly changed.Among the apoptosis related genes,8 pro-apoptotic genes were significantly up-regulated (＞ 6 folds),and them mainly consisted TNFR/TRFSR family (TNFSF8,TNFRSF10B,TNFRSF11 B,TNFRSF9),CIDE family (DFFA),CASP family (CASP10,CASP8),death domain gene family (DAPK1),and the increase of DAPK1 was the most significant (42.83 folds).Three anti-apoptotic genes (CD27,BCL2L10,BIRC4) were significantly upregulated (＞ 6 folds),and 3 genes (BCL2,BAD,BAG) were slightly up-regulated (＞ 2 folds),1 gene (BCL2L1) was slightly down-regulated (＞ 2 folds).Among the autophagy related genes,3 pro-autophagy genes were significantly up-regulated (＞6 folds),4 genes (TNFRSF10A,FADD,TP53,TP53BP2) were slightly up-regulated (＞ 2 folds) ; 3 anti-autophagy genes (BCL2,CASP8,FAS) were slightly up-regulated (＞2 folds),1 gene (MCL1) was slightly down-regulated (＞2 folds).Conclusions ARHI significantly upregulates autophagy-related important regulatory genes Caspse8 and DAPK1.