Detection and Identification of Serum Peptides Biomarker in Papillary Thyroid Cancer

Abstract

BACKGROUND Papillary thyroid cancer (PTC) is currently the most commonly diagnosed endocrine malignancy. In addition, the sex- and age-adjusted incidence of PTC has exhibited a greater increase over the last 2 decades than in many other malignancies. Thus, discovering noninvasive specific serum biomarker to distinguish PTC from cancer-free controls in its early stages remains an important goal. MATERIAL AND METHODS Serum samples from 88 PTC patients and 80 cancer-free controls were randomly allocated into training or validation sets. Serum peptide profiling was performed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS) after using weak cation exchange magnetic beads (WCX-MB), and the results were evaluated by use of ClinProTools™ Software. To distinguish PTC from cancer-free controls, quick classifier (QC), supervised neural network (SNN), and genetic algorithm (GA) models were established. The models were blindly validated to verify their diagnostic capabilities. The most discriminative peaks were subsequently identified with a nano-liquid chromatography-electrospray ionization-tandem mass spectrometry system. RESULTS Six peptide ions were identified as the most discriminative peaks between the PTC and cancer-free control samples. The QC model exhibited satisfactory sensitivity and specificity among the 3 models that were validated. Two peaks, at m/z 2671.17 and m/z 1464.68, were identified as fragments of the alpha chain of fibrinogen, while a peak at m/z 1738.92 was a fragment of complement component 4A/B. CONCLUSIONS MS combined with ClinProTools™ software was able to detect peptide biomarkers in PTC patients. In addition, the constructed classification models provided a serum peptidome pattern for distinguishing PTC from cancer-free controls. Both fibrinogen a and complement C4A/B were identified as potential markers for diagnosis of PTC.

Figure 1

A flowchart of this study. PTC – papillary thyroid cancer; BTN – benign thyroid node; HI – healthy individuals.

Figure 2

ROC curve of peak m/z 1738.92.

Figure 3

Identification of serum peptides present at higher levels in PTC patients. Sequencing of select serum peptides was achieved with LTQ-Orbitrap-MS/MS. (A) Peak m/z 2671.17 (sequence: K.SYKMADEAGSEADHEGTHSTKRGHA.K) was identified as a fragment of fibrinogen alpha chain with an ion score of 196. (B) Peak m/z 1738.92 (sequence: R.NGFKSHALQLNNRQI.R) was identified as a fragment of Complement C4B with an ion score of 64.