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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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
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author Cybulski, Jonathan D.
Leung, Kit Sum
Leung, Carmen Oi Ning
Baker, David M.
Lee, Terence Kin Wah
author_facet Cybulski, Jonathan D.
Leung, Kit Sum
Leung, Carmen Oi Ning
Baker, David M.
Lee, Terence Kin Wah
author_sort Cybulski, Jonathan D.
collection PubMed
description 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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spelling pubmed-104628782023-08-30 Protocol to track the biosynthesis of cholesterol in cultured HCC cells using (13)C compound-specific stable isotopic tracers Cybulski, Jonathan D. Leung, Kit Sum Leung, Carmen Oi Ning Baker, David M. Lee, Terence Kin Wah STAR Protoc Protocol 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. Elsevier 2023-08-17 /pmc/articles/PMC10462878/ /pubmed/37594893 http://dx.doi.org/10.1016/j.xpro.2023.102506 Text en © 2023 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Cybulski, Jonathan D.
Leung, Kit Sum
Leung, Carmen Oi Ning
Baker, David M.
Lee, Terence Kin Wah
Protocol to track the biosynthesis of cholesterol in cultured HCC cells using (13)C compound-specific stable isotopic tracers
title Protocol to track the biosynthesis of cholesterol in cultured HCC cells using (13)C compound-specific stable isotopic tracers
title_full Protocol to track the biosynthesis of cholesterol in cultured HCC cells using (13)C compound-specific stable isotopic tracers
title_fullStr Protocol to track the biosynthesis of cholesterol in cultured HCC cells using (13)C compound-specific stable isotopic tracers
title_full_unstemmed Protocol to track the biosynthesis of cholesterol in cultured HCC cells using (13)C compound-specific stable isotopic tracers
title_short Protocol to track the biosynthesis of cholesterol in cultured HCC cells using (13)C compound-specific stable isotopic tracers
title_sort protocol to track the biosynthesis of cholesterol in cultured hcc cells using (13)c compound-specific stable isotopic tracers
topic Protocol
url 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
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