Abstract:

The effects of overexpression and knockdown of Tmem176a on lipid metabolic pathways in Hepa1-6 cells were investigated.The constructed Tmem176a overexpression lentivirus vector and siRNA were transfected into Hepa1-6 cells. The overexpressed or knocked down with Tmem176a of Hepa1-6 cells were subjected to RNA-seq, bioinformatics analysis, and Real-Time PCR verification.Compared with the control group, Tmem176a was overexpressed 182 times (P<0. 05) and knocked down 0. 2 times (P<0. 05) in Hepa1-6 cells. There were 424 differentially expressed genes (DEGs) in OE-Tmem176a compared with that in OE-Vector, of which 233genes wereupregulated and 192 genes were downregulated. There were 405 DEGs of KD-Tmem176a compared with that of KD-Scramble,of which 157 genes were upregulated and 248 genes were downregulated. Some of the DEGs were enriched in metabolic pathwayssuch as fat digestion and absorption, cholesterol metabolism, glyceride metabolism, and fatty acid synthesis. The Real-Time PCR wasused to investigate some of the DEGs, and the change trend of the expression level of some investigated DEGs was consistent with that of RNAseq. RNAseq results indicated that Tmem176a affected gene response to lipid metabolism, and the changed expression level of Tmem176a promoted significant changes of gene expression level of lipid metabolism in mouse hepatoma Hepa1-6 cells.

Key words: Hepa1-6 cell, lipid meTableolism, RNA-seq, Real-Time PCR