Protocol to track the biosynthesis of cholesterol in cultured HCC cells using (13)C compound-specific stable isotopic tracers

Cholesterol biosynthesis supports proliferation and drives resistance to tyrosine kinase inhibitor (TKI) therapy in hepatocellular carcinoma (HCC). Here, we present a protocol for using stable isotopic tracers to track the biosynthesis of cholesterol in cultured HCC cells. We describe steps for cell...

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Detalles Bibliográficos
Autores principales: Cybulski, Jonathan D., Leung, Kit Sum, Leung, Carmen Oi Ning, Baker, David M., Lee, Terence Kin Wah
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10462878/
https://www.ncbi.nlm.nih.gov/pubmed/37594893
http://dx.doi.org/10.1016/j.xpro.2023.102506
Descripción
Sumario:Cholesterol biosynthesis supports proliferation and drives resistance to tyrosine kinase inhibitor (TKI) therapy in hepatocellular carcinoma (HCC). Here, we present a protocol for using stable isotopic tracers to track the biosynthesis of cholesterol in cultured HCC cells. We describe steps for cell preparation, incubation, separation, and homogenization. We then detail lipid extraction and compound-specific isotope analysis for comparing and quantifying cholesterol synthesis between TKI-resistant HCC cells and their mock counterparts. This protocol can be expanded for use with other shorter-chained lipids.

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