Luciferase does not alter metabolism in cancer cells

Abstract

Luciferase transfected cell lines are used extensively for cancer models, revealing valuable biological information about disease mechanisms. However, these genetically encoded reporters, while useful for monitoring tumor response in cancer models, can impact cell metabolism. Indeed firefly luciferase and fatty acyl-CoA synthetases differ by a single amino acid, raising the possibility that luciferase activity might alter metabolism and introduce experimental artifacts. Therefore knowledge of the metabolic response to luciferase transfection is of significant importance, especially given the thousands of research studies using luciferase as an in vivo bioluminescence imaging (BLI) reporter. Untargeted metabolomics experiments were performed to examine three different types of lymphoblastic leukemia cell lines (Ramos, Raji and SUP T1) commonly used in cancer research, each were analyzed with and without vector transduction. The Raji model was also tested under perturbed starvation conditions to examine potential luciferase-mediated stress responses. The results showed that no significant metabolic differences were observed between parental and luciferase transduced cells for each cell line, and that luciferase overexpression does not alter cell metabolism under basal or perturbed conditions.

Steroid receptor coactivator-3 inhibition generates breast cancer antitumor immune microenvironment

“…Luciferase-labeled E0771 cells were generated by a lentivirus carrying a luciferase expression cassette to noninvasively evaluate tumor growth in mice. Since luciferase expression does not alter metabolism in cancer cells [ 19 ], the luciferase activity measured from luciferase-expressing E0771 breast tumors facilitated the practical quantification of tumor development in recipient mice. As shown in our previous study, continuous treatment of mice with SI-2 (2 mg/kg, twice a day, for 5 weeks) suppressed MDA-MB-468 breast cancer progression in severe combined immunodeficiency (SCID) mice without inducing toxicities in internal organs [ 11 ].…”

Pancreatic cancer growth using magnetic resonance and bioluminescence imaging

“…However, the possibility that luciferase-expression is associated with altered cell growth in vivo has been investigated in the literature with contradictory results [24]. In the present work this point has not been investigated because the focus was the comparison of two imaging modalities using a single cell line.…”

Interactive XCMS Online: Simplifying Advanced Metabolomic Data Processing and Subsequent Statistical Analyses

“…Cell extractions and analyses were performed as described before. 16 For normalization, the cells were counted using the Guava Viacount assay Millipore, Billerica, MA, USA) and placed into 5 mL aliquots of 10 million cells per replicate before extraction, to ensure uniform cell numbers in each replicate.…”