BCAT2 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human hepatic adenocarcinoma cell line SK-HEP-1, designed for the disruption of the BCAT2 gene. This product provides a heterogeneous pool of BCAT2-deficient cells for studying loss-of-function effects in a genetically diverse setting relevant to tumor heterogeneity. This format preserves cellular heterogeneity, mirroring tumor diversity in vivo.
SK-HEP-1 is an epithelial cell line originating from a liver adenocarcinoma, widely used as a hepatocellular carcinoma (HCC) model due to its tumorigenic and metabolic characteristics. The liver??s central role in amino acid metabolism makes this cell line an appropriate host for examining the consequences of BCAT2 knockout on BCAA utilization and metabolic adaptation in hepatic cancer.
BCAT2 encodes the mitochondrial branched-chain amino acid aminotransferase, which catalyzes the reversible transamination of leucine, isoleucine, and valine to their corresponding ??-keto acids, feeding into the TCA cycle via acetyl-CoA and succinyl-CoA generation. BCAT2 expression is regulated by PPARA, PPARGC1A, mTORC1, and amino acid deprivation. The enzyme functions as a homodimer and interacts with the BCKDH complex (BCKDHA, BCKDHB, DBT), linking BCAA catabolism to downstream metabolic and signaling networks, particularly mTORC1-mediated growth control.
In SK-HEP-1 cells, BCAT2 knockout abrogates mitochondrial BCAA transamination, causing accumulation of leucine, isoleucine, and valine and depletion of ??-ketoisocaproate, ??-keto-??-methylvalerate, and ??-ketoisovalerate. This metabolic blockade impairs TCA cycle anaplerosis, reduces mTORC1 signaling (as assessed by phospho-S6K1 downregulation), and suppresses cell proliferation while altering metabolic flexibility. These findings underscore the importance of BCAT2 in sustaining the anabolic and bioenergetic demands of hepatocellular carcinoma and model key aspects of BCAA metabolic disorders. Moreover, these metabolic disruptions parallel the pathophysiology of hypervalinemia and hyperleucinemia.
This knockout polyclonal cell population is a versatile tool for cancer metabolism and BCAA research, enabling investigations into HCC metabolic reprogramming, mTOR pathway dynamics, and drug target validation. Applications include LC-MS quantification of BCAA/BCKA, Seahorse respirometry, cell proliferation and wound healing assays, and flow cytometry. These cells are particularly suited for mechanistic studies linking BCAA metabolism to cell growth and survival. To obtain more information, please contact Ascent Research.
