Deciphering novel biomarkers of lymph node metastasis of thyroid papillary microcarcinoma using proteomic analysis of ultrasound-guided fine-needle aspiration biopsy samples

Abstract

Thyroid papillary microcarcinoma is now a common clinical problem. Cervical lymph node metastasis is the main metastasis mode of PTMC. However, before operation, it is still difficult to determine exactly whether PTMC patient is suffering with cervical lymph node metastasis. To resolve this dilemma, for better selection of optimum treatment plans, it is necessary to investigate the overall changes in proteomes of PTMC, and evaluate the potential of biomarkers to predict lymph node metastasis. Tandem mass tags combined with multidimensional liquid chromatography and mass spectrometry analyses were used aiming to screen the proteomic profiles of fine-needle aspiration biopsy samples. Quantitative proteomic analysis, significant pathway and functional categories were investigated. In total, 3391 proteins of the 3793 protein groups identified were quantified. Bioinformatics analysis indicated that differentially expressed proteins were involved in multiple biological functions, metastasis-related pathways. Moreover, IFN-stimulated gene 15 proteins were found to be well distinguished between patients with lymph node metastatic and patients with nonmetastatic PTMC. Knocking down ISG15 with shRNA inhibited the xenografted tumor growth. This study provided a reference proteome map for lymph node metastatic PTMC. ISG15 probably is a prognosis marker of thyroid papillary microcarcinoma patients with lymph node metastasis. SIGNIFICANCE: Nowadays, thyroid cancer has become a widespread epidemic. The rate of thyroid cancer incidence has been faster than any other cancers, reported by the American Cancer Society. Papillary thyroid microcarcinoma (PTMC) is a subset of PTC defined as PTC measuring≤1 cm in size, which comprises nearly one-half of all the cases of PTCs. Actually, the rapidly increasing global incidence of PTC is mainly attributed to the corresponding increase in the diagnosis of PTMC. Scholars have figuratively compared the increase of PTMC to the "tsunami". The treatment scheme for PTMC is still not uniform, and the controversy is mainly focused on the necessity of surgery treatment. PTMCs often have an indolent course in the absence of evidence of metastatic cervical lymph nodes, distant metastases and extrathyroidal extension. Therefore, it is important for us to reliably differentiate the small number of PTMC patients developing significant metastases progression from the larger population of patients that harbor indolent PTMCs. The present study aimed to investigate the overall changes in proteomes of PTMC, and evaluate the potential of biomarkers to predict lymph node metastasis. Tandem mass tags (TMT) combined with multidimensional liquid chromatography and mass spectrometry analyses were used aiming to screen the proteomic profiles of fine-needle aspiration biopsy (FNAB) samples. Quantitative proteomic analysis, significant pathway and functional categories were investigated. Our results showed that some differential expression proteins were likely to be important resources for finding new diagnostic biomarkers.

Keywords: Fine needle aspiration biopsy; Lymph node metastasis;biomarker; Papillary microcarcinoma; Proteomics.