RNA Sequencing of RNA profiles from BHMT Overexpression (OE) and BHMT knockout (KO)

Purpose: Betaine-Homocysteine Methyltransferase (BHMT) catalyzes the conversion of betaine to dimethylglycine and homocysteine to methionine. We were interested in both overexpressing and knocking out BHMT and examining the global effects of RNA transcription. We did this in both in vivo and in vitro models. Methods: Tumors: 1,000,000 HepG2 BHMT OE or BHMT KO cells were injected to the left and right flank of nude mice (n = 5), respectively, as a xenograft. Once tumors reached the appropriate size, they were harvested. The tissue was homogenized and RNA was extracted. Cells: HepG2 BHMT OE and KO cells (n = 3/group) were grown in Human Plasma-Like Medium (HPLM) for 48 hours. Then, cells were harvested and RNA was extracted. Both: Samples were submitted to the University of Utah sequencing core, who performed library prep and sequencing. Total RNA was taken from samples and run on an illumina sequencer. Note: OE2 and KO2 were large outliers, and therefore remove from the data analysis. They came from the same mouse.