Simultaneous Determination of Methylated Nucleosides by HILIC-MS/MS Revealed Their Alterations in Urine from Breast Cancer Patients.
Zhihao FangYiqiu HuXiujuan HongXiaoxiao ZhangTao PanChi PanShu ZhengCheng GuoPublished in: Metabolites (2022)
RNA methylation plays a vital role in the pathogenesis of a variety of diseases including cancer, and aberrant levels of modified nucleosides in RNA were revealed to be related to cancer. Urine is a favored source for biomarker discovery due to the non-invasion to patients. Herein, we developed a sensitive hydrophilic interaction liquid chromatography tandem mass spectrometry (HILIC-MS/MS) method combined with stable isotope dilution for accurate quantification of methylated nucleosides in human urine. With this method, we successfully quantified ten methylated nucleosides in urine samples collected from healthy controls and breast cancer patients. We found N 6 -methyladenosine (m 6 A), 2'-O-methyladenosine (A m ), N 1 -methyladenosine (m 1 A), N 6 ,2'-O-dimethyladenosine (m 6 A m ), N 1 -methylguanosine (m 1 G), 2'-O-methylguanosine (G m ), 5-methylcytidine (m 5 C) and 2'-O-methylcytidine (C m ) were all decreased in early-stage breast cancer patients, and a nomogram prediction model was constructed. Locally advanced breast cancer patients exhibited elevated levels of urinary 2'-O-methylated nucleosides in comparison to early-stage breast cancer patients. Together, we developed a robust method for the simultaneous determination of methylated nucleosides in human urine, and the results revealed an association between the contents of urinary methylated nucleosides and the occurrence of breast cancer, which may stimulate future studies about the regulatory roles of these methylated nucleosides in the initiation and progression of breast cancer.
Keyphrases
- liquid chromatography tandem mass spectrometry
- simultaneous determination
- ms ms
- early stage
- solid phase extraction
- liquid chromatography
- high performance liquid chromatography
- tandem mass spectrometry
- ultra high performance liquid chromatography
- endothelial cells
- locally advanced
- end stage renal disease
- single cell
- papillary thyroid
- chronic kidney disease
- mass spectrometry
- squamous cell carcinoma
- small molecule
- ejection fraction
- high resolution mass spectrometry
- induced pluripotent stem cells
- dna methylation
- neoadjuvant chemotherapy
- prognostic factors
- radiation therapy
- risk assessment
- wastewater treatment
- cell migration
- high resolution
- pluripotent stem cells
- young adults
- gas chromatography
- current status