Expression data from hepatocellular carcinoma cell lines

In this study, we demonstrated that mitochondrial respiratory defect enhanced NFE2L1 transcription via reactive oxygen species (ROS)-mediated STAT3 activation and the up-expressed NFE2L1 increased hepatoma cell invasiveness by inducing syntaxin 12 (STX12) expression. Bioinformatic analysis of The Cancer Genome Atlas Liver Hepatocellular carcinoma (TCGA-LIHC) open database showed that NFE2L1 expression is strongly positively correlated with STX12 expression and, furthermore, epithelial-mesenchymal transition (EMT)-related core genes were significantly upregulated in the tumors expressing both NFE2L1 and STX12. The effect of NFE2L1/STX12 axis on lung metastasis of hepatoma cell was proved in nude mice model. Collectively, our results indicate that NFE2L1 was a key mitochondrial retrograde signaling-mediated primary gene product to enhance hepatoma cell invasiveness via STX12 expression To identify for downstream effector gene of NFE2L1, we depleted NFE2L1 in SNU423 and overexpressed it in SNU387. For this, cells were transfected with the plasmids and target siRNA duplexes using FuGENE® HD (Promega) and OligofectamineTM Reagent (Invitrogen), respectively, according to the manufacturer’s instructions. For transfection, we used two siRNA targeting different sequence of NFE2L1 to deplete the NFE2L1 expression in SNU423 and one plasmid to overexpress the NFE2L1 expression in SNU387. Each transfection was duplicated and total 10 samples were used for microarry gene expression profiling.