Recombinant pEGFP‑N1‑IRF3a Plasmid Induces the Apoptosis of Non-Small Cell Lung Cancer Cells by Regulating the Activity of STAT1
YI Liang1, WU Yanping2, HAN Qian2, YANG Yunmei1*
IRF3 (interferon regulatory factor 3) plays the significant roles in regulating innate immune activity. IRF3a (interferon regulatory factor 3 isoform 3) is the translated production of IRF3 transcript variant 3 under the control of alternative splicing. However, seldom researches whether IRF3a has effect on the apoptosis of cancer cells been performed. In this research, the level of IRF3a gene expression was firstly tested in the lung cancer tissues and the adjacent tissues by qRT-PCR. IRF3a gene was obtained from human peripheral blood mononuclear cells by PCR and cloned into pEGFP-N1 plasmid, then the recombinant pEGFP-N1-IRF3a plasmid was successfully constructed. The apoptotic cells dramatically increased and the level of cleaved caspase3, cleaved caspase8 strikingly elevated followed recombinant pEGFP-N1-IRF3a plasmid transfected into A549 and H1299 cells. Moreover, IRF3a overexpression could promote the activity of STAT1 and the apoptotic cells decreased obviously following the pretreatment of STAT1 inhibitor Nifuroxazide. Therefore, recombinant pEGFP-N1-IRF3a plasmid was successfully constructed in this research and it further revealed that IRF3a promoted the apoptosis of NSCLC A549 and H1299 cells by activating the phosphorylation of STAT1.
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