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利用荧光衍生化-高效液相色谱法通过氨基酸代谢组学评估抗癌药物的疗效

Assessment of the efficacy of anticancer drugs by amino acid metabolomics using fluorescence derivatization-HPLC.

作者信息

Tomita Ryoko, Todoroki Kenichiro, Machida Kazuyuki, Nishida Sho, Maruoka Hiroshi, Yoshida Hideyuki, Fujioka Toshihiro, Nakashima Manabu, Yamaguchi Masatoshi, Nohta Hitoshi

机构信息

Faculty of Pharmaceutical Sciences, Fukuoka University.

出版信息

Anal Sci. 2014;30(7):751-8. doi: 10.2116/analsci.30.751.

Abstract

Metabolomic studies conducted for evaluating cancer pathogenesis and progression by monitoring the amino acids metabolic balance hold great promise for assessing current and future anticancer treatments. We performed a comprehensive quantification of 21 amino acids concentrations in cultured human colorectal adenocarcinoma cells treated with the anticancer drugs 5-fluorouracil, irinotecan, and cisplatin. A precolumn fluorescence derivatization-HPLC method involving 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate was used. Amino acid concentration data were analyzed by principal-component analysis and partial least-squares multivariate statistical methods to represent samples on two-dimensional graphs. The hierarchical cluster analysis and linear discriminant analysis were used to classify the samples on the score plots. Unlike the cluster analysis approach, the linear discrimination analysis classification successfully distinguished anticancer drug-treated samples from the untreated controls. Moreover, three candidate amino acids (serine, aspartic acid, and methionine) were identified from the loading plots as potential biomarkers. Our proposed method might be able to evaluate the effectiveness of anticancer therapy even in small laboratories or medical institutions.

摘要

通过监测氨基酸代谢平衡来评估癌症发病机制和进展的代谢组学研究,在评估当前和未来的抗癌治疗方面具有巨大潜力。我们对用抗癌药物5-氟尿嘧啶、伊立替康和顺铂处理的培养人结肠直肠腺癌细胞中的21种氨基酸浓度进行了全面定量。使用了一种涉及6-氨基喹啉-N-羟基琥珀酰亚胺基氨基甲酸酯的柱前荧光衍生化-高效液相色谱法。通过主成分分析和偏最小二乘多元统计方法分析氨基酸浓度数据,以在二维图上表示样本。层次聚类分析和线性判别分析用于在得分图上对样本进行分类。与聚类分析方法不同,线性判别分析分类成功地将抗癌药物处理的样本与未处理的对照区分开来。此外,从载荷图中鉴定出三种候选氨基酸(丝氨酸、天冬氨酸和蛋氨酸)作为潜在生物标志物。我们提出的方法即使在小型实验室或医疗机构中也可能能够评估抗癌治疗的有效性。

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