Protocol to track the biosynthesis of cholesterol in cultured HCC cells using 13C compound-specific stable isotopic tracers
Summary: 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...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-09-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723004732 |
| _version_ | 1828716942122287104 |
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| author | Jonathan D. Cybulski Kit Sum Leung Carmen Oi Ning Leung David M. Baker Terence Kin Wah Lee |
| author_facet | Jonathan D. Cybulski Kit Sum Leung Carmen Oi Ning Leung David M. Baker Terence Kin Wah Lee |
| author_sort | Jonathan D. Cybulski |
| collection | DOAJ |
| description | Summary: 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. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-12T14:20:41Z |
| format | Article |
| id | doaj.art-aece1a8784524daf96df78805392605a |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-12T14:20:41Z |
| publishDate | 2023-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-aece1a8784524daf96df78805392605a2023-08-19T04:32:34ZengElsevierSTAR Protocols2666-16672023-09-0143102506Protocol to track the biosynthesis of cholesterol in cultured HCC cells using 13C compound-specific stable isotopic tracersJonathan D. Cybulski0Kit Sum Leung1Carmen Oi Ning Leung2David M. Baker3Terence Kin Wah Lee4School of Biological Sciences, Faculty of Science, The University of Hong Kong, Hong Kong, Hong Kong SAR; Smithsonian Tropical Research Institute, Balboa, Republic of Panama; Corresponding authorSmithsonian Tropical Research Institute, Balboa, Republic of PanamaDepartment of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hong Kong, Hong Kong SARSmithsonian Tropical Research Institute, Balboa, Republic of Panama; Corresponding authorDepartment of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hong Kong, Hong Kong SAR; State Key Laboratory of Chemical Biology and Drug Discovery, The Hong Kong Polytechnic University, Hong Kong, Hong Kong SAR; Corresponding authorSummary: 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. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166723004732Cell CultureCancerMetabolismChemistry |
| spellingShingle | Jonathan D. Cybulski Kit Sum Leung Carmen Oi Ning Leung David M. Baker Terence Kin Wah Lee Protocol to track the biosynthesis of cholesterol in cultured HCC cells using 13C compound-specific stable isotopic tracers STAR Protocols Cell Culture Cancer Metabolism Chemistry |
| title | Protocol to track the biosynthesis of cholesterol in cultured HCC cells using 13C compound-specific stable isotopic tracers |
| title_full | Protocol to track the biosynthesis of cholesterol in cultured HCC cells using 13C compound-specific stable isotopic tracers |
| title_fullStr | Protocol to track the biosynthesis of cholesterol in cultured HCC cells using 13C compound-specific stable isotopic tracers |
| title_full_unstemmed | Protocol to track the biosynthesis of cholesterol in cultured HCC cells using 13C compound-specific stable isotopic tracers |
| title_short | Protocol to track the biosynthesis of cholesterol in cultured HCC cells using 13C compound-specific stable isotopic tracers |
| title_sort | protocol to track the biosynthesis of cholesterol in cultured hcc cells using 13c compound specific stable isotopic tracers |
| topic | Cell Culture Cancer Metabolism Chemistry |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723004732 |
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