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首页> 外文期刊>Artificial cells, blood substitutes, and immobilization biotechnology. >Synergistic Effect of IFN-γ Gene on LIGHT-induced Apoptosis in HepG2 Cells via Down Regulation of Bcl-2
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Synergistic Effect of IFN-γ Gene on LIGHT-induced Apoptosis in HepG2 Cells via Down Regulation of Bcl-2

机译：IFN-γ基因通过下调Bcl-2对光诱导HepG2细胞凋亡的协同作用

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: To detect the expression of anti-apoptotic factor Bcl-2 and Survivin in transferred HepG2 cells and evaluate the synergistic effect of IFN-γ gene on LIGHT-induced apoptosis signal transduction pathways, the full-length ORF of LIGHT and IFN-γ gene were cloned into pcDNA4 and verified by DNA sequencing. After being optimized by EGFP, recombinant LIGHT and IFN-γ were transferred into the HepG2 cells mediated by a cationic liposome in vitro. The expression of LIGHT and IFN-γ was identified in the supernatants by ELISA. The HepG2 cells were divided into three groups: the control, LIGHT gene transfection alone, and simultaneous transfection of LIGHT and IFN-γ genes. The cell apoptosis and expression of Bcl-2 and Survivin in cell lysate were detected through FCM. After transfection, the apoptosis rate of HepG2 cells was increased with the prolonged time, and the apoptosis rate of LIGHT group was higher than thecontrol group, while the LIGHT/IFN-γ group was higher than the LIGHT group P 0.01). The expression of Bcl-2 and Survivin in LIGHT group and LIGHT/IFN-γ group decreased dramatically compared with the control group. LIGHT gene alone can result in significant inhibition of HepG2 cells proliferation. INF-γ can synergistically precede LIGHT-induced apoptotic processes through down-regulation of Bcl-2 expression, but not survivin expression.
机译：：检测抗凋亡因子Bcl-2和Survivin在转移的HepG2细胞中的表达，并评估IFN-γ基因对LIGHT诱导的细胞凋亡信号转导通路，LIGHT的全长ORF和IFN-γ基因的协同作用将其克隆到pcDNA4中并通过DNA测序验证。经EGFP优化后，重组LIGHT和IFN-γ在体外被阳离子脂质体介导转移到HepG2细胞中。通过ELISA鉴定上清液中的LIGHT和IFN-γ的表达。 HepG2细胞分为三组：对照组，单独的LIGHT基因转染以及同时转染LIGHT和IFN-γ基因。通过FCM检测细胞裂解液中的细胞凋亡以及Bcl-2和Survivin的表达。转染后，HepG2细胞的凋亡率随时间的延长而增加，LIGHT组的细胞凋亡率高于对照组，而LIGHT /IFN-γ组的细胞凋亡率高于LIGHT组（P <0.01）。与对照组相比，LIGHT组和LIGHT /IFN-γ组Bcl-2和Survivin的表达明显降低。单独的LIGHT基因可以显着抑制HepG2细胞的增殖。 INF-γ可以通过下调Bcl-2表达而不是survivin表达来协同先于LIGHT诱导的凋亡过程。

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