亲水作用色谱/质谱联用方法用于膀胱癌患者血清代谢组学研究

Hydrophilic Interaction Liquid Chromatography Coupled with Mass Spectrometry for Serum Metabolomics Analysis of Bladder Cancer

膀胱癌是泌尿系统最常见的恶性肿瘤之一,具有高发病率、高复发率和高进展率的特点。本研究应用69个极性代谢物标样选择合适的分离系统,建立了两性离子亲水作用色谱/质谱联用的代谢组学分析方法。本方法线性范围较宽,检出限低于ng/m L数量级。将本方法用于血清代谢组学分析,85%以上代谢物峰面积的RSD<30%。对64例膀胱癌患者和32例正常人的血清进行代谢组学研究,发现溶血磷脂酰胆碱、游离脂肪酸、氨基酸、胆汁酸、有机酸、核苷等在患病组和正常组中存在显著差异。经筛选和验证,甘磷酸胆碱、胱氨酸、十二碳烯酸、二十碳烯酸和鹅去氧胆酸5种代谢物可以作为区分膀胱癌和正常人的潜在标志物。本研究结果表明,基于亲水作用色谱/质谱联用的代谢组学方法是发现癌症诊断潜在生物标志物的有效手段。

Bladder cancer( BC) is a fatal malignancy with considerable mortality,and can cause a serious threat to human health. The successful treatment of bladder cancer relies mainly on early detection. Biomarkers are vital to early diagnosis of bladder cancer,and metabonomics play an important role in biomarkers finding.In this study,we used 69 polar metabolites to select the appropriate separation system and develop the zwitterionic hydrophilic chromatography/mass spectrometry( ZIC-HILIC/MS) method. In this method,50 representative compounds had broad linear ranges between 2-6 orders of magnitude. Moreover the limit of detection of the method was below ng/m L levels. The analysis for six serum samples prepared in parallel showed that this method had good reproducibility,and the RSDs of more than 85% metabolites were less than30%. Based on this method,it was found that 35 metabolites had significant differences in BC group and healthy control. After screening and validation,the combination of chenodeoxycholic acid,eicosenoic acid,GPC,dodecenoic acid and cystine was a potential biomarker to distinguish BC and normal group. These results indicated that the ZIC-HILIC/MS method could detect diverse metabolites for metabolomic analysis purpose with good reproducibility and stability.