Abstract
DNA damage induced apoptosis suppressor (DDIAS), or human Noxin (hNoxin), is strongly expressed in lung cancers. DDIAS knockdown induced apoptosis in non-small cell lung carcinoma A549 cells in response to DNA damage, indicating DDIAS as a potential therapeutic target in lung cancer. To understand the transcriptional regulation of DDIAS, we determined the transcription start site, promoter region, and transcription factor. We found that DDIAS transcription begins at nucleotide 212 upstream of the DDIAS translation start site. We cloned the DDIAS promoter region and identified NFAT2 as a major transcription factor (Im et al., 2016 [1]). We demonstrated that NFATc1 regulates DDIAS expression in both pancreatic cancer Panc-1 cells and lung cancer cells.
Highlights
DNA damage induced apoptosis suppressor (DDIAS), or human Noxin, is strongly expressed in lung cancers
We found that DDIAS transcription begins at nucleotide 212 upstream of the DDIAS translation start site
We demonstrated that NFATc1 regulates DDIAS expression in both pancreatic cancer Panc-1 cells and lung cancer cells
Summary
HEK293, non-small cell lung cancer cell NCI-H1703, and pancreatic cancer cell Panc-1 cells were cultured in an incubator at 37 °C and 5% CO2. HEK293 and Panc-1 cells were cultured in Dulbecco's modified Eagle's medium (DMEM); NCI-H1703 cells were cultured in RPMI-1640 containing 10% fetal bovine serum (FBS), 50 U/mL of penicillin, and 50 μg/mL of streptomycin (Invitrogen, USA). 40 nM siRNAs with Lipofectamine 2000 (Invitrogen, USA) according to the manufacturer's instructions. SiRNA sequences were as follows: siSP1, 50-CUUUUUCACCAAUGCCAAU-30, sicJun: 50-CAAGAAGAUGCGCC GCAAC-30, sicMyc: 50-GUCAAGAGGCGAACACACA-30, siNFATc1: 50-GGACUCCAAGGUCAUUUUC-30, siDDIAS D1: 50-GCCUAUCAUUUCCCUGAUC-30, siDDIAS D2: 50-CUGUAACCCAGGCAGAUGU-30, or siScrambled: 50CCUACGCCACCAAUUUCGU-30 40 nM siRNAs with Lipofectamine 2000 (Invitrogen, USA) according to the manufacturer's instructions. siRNA sequences were as follows: siSP1, 50-CUUUUUCACCAAUGCCAAU-30, sicJun: 50-CAAGAAGAUGCGCC GCAAC-30, sicMyc: 50-GUCAAGAGGCGAACACACA-30, siNFATc1: 50-GGACUCCAAGGUCAUUUUC-30, siDDIAS D1: 50-GCCUAUCAUUUCCCUGAUC-30, siDDIAS D2: 50-CUGUAACCCAGGCAGAUGU-30, or siScrambled: 50CCUACGCCACCAAUUUCGU-30
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