Method for Lipidomic Analysis: p53 Expression Modulation of Sulfatide, Ganglioside, and Phospholipid Composition of U87 MG Glioblastoma Cells
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- 作者：Huan He ; Charles A. Conrad ; Carol L. Nilsson ; Yongjie Ji ; Tanner M. Schaub ; Alan G. Marshall ; Mark R. Emmett
- 刊名：Analytical Chemistry
- 出版年：2007
- 出版时间：November 15, 2007
- 年：2007
- 卷：79
- 期：22
- 页码：8423 - 8430
- 全文大小：260K
- 年卷期：v.79,no.22(November 15, 2007)
- ISSN：1520-6882
文摘
Lipidomics can complement genomics and proteomics byproviding new insight into dynamic changes in biomembranes; however, few reports in the literature have explored, on an organism-wide scale, the functional linkbetween nonenzymatic proteins and cellular lipids. Here,we report changes induced by adenovirus-delivered wild-type p53 gene and chemotherapy of U87 MG glioblastomacells, a treatment known to trigger apoptosis and cell cyclearrest. We compare polar lipid changes in treated cellsand control cells by use of a novel, sensitive method thatemploys lipid extraction, one-step liquid chromatographyseparation, high-resolution mass analysis, and Kendrickmass defect analysis. Nano-LC FT-ICR MS and quadrupole linear ion trap MS/MS analysis of polar lipids yieldshundreds of unique assignments of glyco- and phospholipids at sub-ppm mass accuracy and high resolving power(m/m50% = 200 000 at m/z 400) at 1 s/scan. MS/MSdata confirm molecular structures in many instances.Sulfatides are most highly modulated by wild-type p53treatment. The treatment also leads to an increase inphospholipids such as phosphatidyl inositols, phosphatidyl serines, phosphatidyl glycerols, and phosphatidylethanolamines. An increase in hydroxylated phospholipids is especially noteworthy. Also, a decrease in the longerchain gangliosides, GD1 and GM1b, is observed in wild-type p53 (treated) cells.