Comprehensive lipidome profiling of isogenic primary and metastatic colon adenocarcinoma cell lines.
Comprehensive lipidome profiling of isogenic primary and metastatic colon adenocarcinoma cell lines.
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DOI：
10.1021/ac302154g
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发表时间：
2012-11-06
影响因子：
7.4
通讯作者：
Reid, Gavin E.
中科院分区：
文献类型：
--
作者：
Fhaner, Cassie J.;Liu, Sichang;Ji, Hong;Simpson, Richard J.;Reid, Gavin E.
一种由连续的官能团选择性化学修饰反应、高分辨率/准确的质谱学和“靶向”串联质谱学(MS/MS)分析组成的“鸟枪式”脂质组学策略已被开发用于全面鉴定、表征和定量分析>；600单个甘油磷脂、甘油脂、鞘磷脂和类固醇脂类的相对丰度的变化，该细胞系原发结直肠癌(CRC)细胞系SW480及其同基因的淋巴转移衍生物SW620。甘油磷乙醇胺和甘油磷酸丝氨酸的选择性化学衍生化使用了含有固定电荷的硫离子d6-S，S‘-二甲硫丁酰羟基丁二酰亚胺酯(d6-DMBNHS)试剂，以消除这些物种与粗提物中所有其他脂肪的前体离子等压质量重叠的可能性，从而能够明确地对它们进行归属，而随后使用甲酸选择性地温和酸解含有脂类的质膜(乙烯基醚)，使得这些物种能够很容易地与含有脂类的等压质量等压浆基醚区分开来。使用这种方法，在SW480和SW620细胞之间观察到了许多先前被认为与癌症进展相关的或在一系列生理和病理过程中发挥中介作用的脂类物种的丰度在统计学上的显著差异。最值得注意的是，与SW480细胞相比，SW620细胞中的血浆胆碱和甘油三酯水平升高，血浆乙醇胺类脂水平降低，含有鞘磷脂和神经酰胺的C-16脂水平降低，总胆固醇酯和甘油三酯类脂的丰度显著增加。
A ‘shotgun’ lipidomics strategy consisting of sequential functional group selective chemical modification reactions coupled with high-resolution / accurate mass spectrometry and ‘targeted’ tandem mass spectrometry (MS/MS) analysis has been developed and applied toward the comprehensive identification, characterization and quantitative analysis of changes in relative abundances of >600 individual glycerophospholipid, glycerolipid, sphingolipid and sterol lipids between a primary colorectal cancer (CRC) cell line, SW480, and its isogenic lymph node metastasized derivative, SW620. Selective chemical derivatization of glycerophosphoethanolamine and glycerophosphoserine lipids using a ‘fixed charge’ sulfonium ion containing, d6-S,S′-dimethylthiobutanoylhydroxysuccinimide ester (d6-DMBNHS) reagent was used to eliminate the possibility of isobaric mass overlap of these species with the precursor ions of all other lipids in the crude extracts, thereby enabling their unambiguous assignment, while subsequent selective mild acid hydrolysis of plasmenyl (vinyl-ether) containing lipids using formic acid enabled these species to be readily differentiated from isobaric mass plasmanyl (alkyl-ether) containing lipids. Using this approach, statistically significant differences in the abundances of numerous lipid species previously identified as being associated with cancer progression, or that play known roles as mediators in a range of physiological and pathological processes, were observed between the SW480 and SW620 cells. Most notably, these included increased plasmanylcholine and triglyceride lipid levels, decreased plasmenylethanolamine lipids, decreased C-16 containing sphingomyelin and ceramide lipid levels, and a dramatic increase in the abundances of total cholesterol ester and triglyceride lipids in the SW620 cells compared to those in the SW480 cells.
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DOI：
10.1016/j.jasms.2005.04.017
发表时间：
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影响因子：
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