基于LC-MS/MS多反应监测技术的食管鳞状细胞癌能量代谢物分析

doi:10.13481/j.1671-587X.20230519

摘要/Abstract

摘要：

目的分析食管鳞状细胞癌（ESCC）患者的血清代谢物成分，筛选出潜在的差异代谢物，探讨ESCC的发生和能量代谢之间的关系，为ESCC的早期诊断提供有价值的能量代谢物。方法收集ESCC患者（ESCC组）和健康志愿者（对照组）的血清样本，每组7例。采用超高效液相色谱串联质谱（LC-MS/MS）技术检测2组研究对象血清代谢物。通过偏最小二乘判别分析（PLS-DA）模型和正交偏最小二乘判别分析（OPLS-DA）模型鉴定ESCC组和对照组研究对象血清代谢物是否存在差异，并按照血清代谢物的化学分类归属进行分类。基于多反应监测（MRM）模式靶向代谢组学分析血清能量代谢物，确定显著差异能量代谢物及其通路。结果纳入ESCC组和对照组血清样本进行检测，正和负离子模式合并后共鉴定到 266 种代谢物，2种模式分别鉴定到164和102种代谢物。单变量统计分析，ESCC组和对照组研究对象血清代谢物存在显著差异。在有化学分类归属的代谢物中，脂质和类脂质分子占比最高（18.421%）。ESCC组中显著性差异能量代谢物共有3种，其中L-苹果酸和异柠檬酸表达水平上调、环磷腺苷（cAMP）表达水平下调。结论 ESCC患者和健康志愿者血清能量代谢物表达存在差异，ESCC的发生与能量代谢有关。

关键词: 食管鳞状细胞肿瘤, 代谢组学, 能量代谢物, 多反应监测模式, 代谢物

Abstract:

Objective： To analyze the serum metabolite compositions of the patients with esophageal squamous cell carcinoma （ESCC）， to screen out the potential differential metabolites， and to explore the relationship between the occurrence of ESCC and energy metabolism， and to provide the useful energy metabolites for the early diagnosis of ESCC. Methods The serum samples of the ESCC patients and healthy volunteers were collected and divided into ESCC group and control group， and there were 7 cases in each group. Liquid chromatography-tandem mass spectrometry （LC-MS/MS） method was used to detect the serum metabolites of the subjects in two groups；partial least squares discriminant analysis （PLS-DA） and orthogonal partial least squares discriminant analysis （OPLS-DA） models were used to identify the differences in serum metabolites of the subjects between ESCC group and control group， and they were classified according to the chemical classification of serum metabolites； targeted metabolomics analysis on serum energy metabolites was conducted based on the multi-response monitoring （MRM） mode to further identify the significantly differential energy metabolites and their pathways. Results A total of 266 metabolites were identified in the serum samples of the subjects in ESCC group and control group with the positive and negative ion modes， and there were 164 and 102 metabolites identified with the above two modes， respectively. The univariate statistical analysis results showed that there were significant differences in serum metabolites of the subjects between ESCC group and control group. Among the metabolites with chemical classification， lipids and lipid-like molecules accounted for the highest proportion （18.421%）. Three significantly different energy metabolites in ESCC group were found， and the expression levels of L-malic acid and isocitric acid were increased and the expression level of cyclic adenosine monophosphate （cAMP） was decreased. Conclusion There are differences in the expressions of serum energy metabolites between the ESCC patients and healthy controls， and the occurrence of ESCC is related to the energy metabolism.

Key words: Esophageal squamous cell neoplasms, Metabolomics, Energy metabolites, Multi-response monitoring, Metabolites

中图分类号:

R735.1

董宇航,高海霞,崔晓宾,李锋. 基于LC-MS/MS多反应监测技术的食管鳞状细胞癌能量代谢物分析[J]. 吉林大学学报(医学版), 2023, 49(5): 1253-1261.

Yuhang DONG,Haixia GAO,Xiaobin CUI,Feng LI. Energy metabolite analysis on ESCC based on LC-MS/MS multi-response monitoring technology[J]. Journal of Jilin University(Medicine Edition), 2023, 49(5): 1253-1261.

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Group	Polarity	A	Q²（cum）
CA vs N	Positive	3	0.768
CA vs N	Negative	2	0.478

Group	Polarity	A	Q²（cum）
CA vs N	Positive	1+2	0.50
CA vs N	Negative	1+2	0.58

Metabolite name	ESCC	Control	Transitions	Retention time(t/min)	RSD	FC	P
cAMP	6 962±3 142	11 773±4 513	328/134	1.946	0.019	0.591	0.037
L-malic acid	4 279 565±1 523 940	2 758 430±887 567	133/115	5.090	0.048	1.551	0.039
Isocitrate	57 661±18 673	38 927±12 876	191/73	7.697	0.086	1.481	0.043
Oxaloacetate	102 305±39 628	136 870±39 968	131/87	4.488	0.015	0.747	0.067
Citrate	5 434 073±2 992 632	3 329 249±1 215 777	191/87	7.767	0.063	1.632	0.114
Cis-aconitate	2 829 596±1 389 602	1 931 888±485 449	173/85	6.952	0.071	1.465	0.132
Lactate	1 725 703±406 487	1 524 237±431 240	89/43	1.796	0.033	1.132	0.256
Alpha-ketoglutarate	1 615 659±1 154 810	1 129 208±419 534	145/101	3.877	0.032	1.431	0.320
Dihydroxyacetone phosphate	2 274 830±201 126	2 361 020±794 818	169/97	6.172	0.090	0.963	0.478
Succinate	1 836 588±465 933	1 746 055±822 155	117/73	4.655	0.055	1.052	0.789
Adenosine monophosphate	6 118±1 875	5 909±2 205	346/79	6.114	0.091	1.035	0.836

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