Quantitative SWATH-Based Proteomic Profiling for Identification of Mechanism-Driven Diagnostic Biomarkers Conferring in the Progression of Metastatic Prostate Cancer.

Anshika N Singh,Neeti Sharma

doi:10.3389/fonc.2020.00493

Abstract

Prostate cancer (PCa), the most frequently diagnosed malignancy in men is associated with significant mortality and morbidity. Therefore, demand exists for the identification of potential biomarkers for patient stratification according to prognostic risks and the mechanisms involved in cancer development and progression to avoid over/under treatment of patients and prevent relapse. Quantitative proteomic mass spectrometry profiling and gene enrichment analysis of TGF-β induced-EMT in human Prostate androgen-dependent (LNCaP) and androgen-independent (PC-3) adenocarcinoma cell lines was performed to investigate proteomics involved in Prostate carcinogenesis and their effect onto the survival of PCa patients. Amongst 1,795 proteins, which were analyzed, 474 proteins were significantly deregulated. These proteins contributed to apoptosis, gluconeogenesis, transcriptional regulation, RNA splicing, cell cycle, and MAPK cascade and hence indicating the crucial roles of these proteins in PCa initiation and progression. We have identified a panel of six proteins viz., GOT1, HNRNPA2B1, MAPK1, PAK2, UBE2N, and YWHAB, which contribute to cancer development, and the transition of PCa from androgen dependent to independent stages. The prognostic values of identified proteins were evaluated using UALCAN, GEPIA, and HPA datasets. The results demonstrate the utility of SWATH-LC-MS/MS for understanding the proteomics involved in EMT transition of PCa and identification of clinically relevant proteomic biomarkers.

Highlights

Prostate cancer (PCa) in men is known to be the most frequently diagnosed noncutaneous solid organ cancer and the second major cause of cancer mortalities in the United States [1]
The expression levels of the characteristic markers of epithelial to mesenchymal transition (EMT) viz., E-cadherin, N-cadherin, and Vimentin were evaluated by real-time polymerase chain reaction (PCR) and Western blot to determine the effectiveness of the TGF-β treatment on EMT markers
The analysis showed that the treatment with TGF-β downregulated the expressions of Ecadherin with significant upregulation of mesenchymal markers (N-cadherin and Vimentin) in both androgen-dependent and androgen-independent PCa cell lines

Summary

Introduction

Prostate cancer (PCa) in men is known to be the most frequently diagnosed noncutaneous solid organ cancer and the second major cause of cancer mortalities in the United States [1]. The prostate tumor is observed as a growing indolent tumor or advanced aggressive cancer according to evidence the currently available diagnostic biomarker. Proteomic Biomarkers for Prostate Cancer prostate-specific antigen (PSA) and histological examinations of tumor tissues cannot completely specify the tumor stages, as well as its aggressiveness [2]. The histological samples of tumor biopsies are graded on the basis of Gleason score [2,3,4,5,6,7,8,9,10] that indicates the aggressiveness of the tumor and its metastatic potential [3]. Gleason score can fail at times due to molecular and clinical heterogeneities of Prostate tumors

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Discussion

Conclusion