A Proteomic Approach for the Identification of Up-Regulated Proteins Involved in the Metabolic Process of the Leiomyoma

Abstract

Uterine leiomyoma is the most common benign smooth muscle cell tumor of the uterus. Proteomics is a powerful tool for the analysis of complex mixtures of proteins. In our study, we focused on proteins that were upregulated in the leiomyoma compared to the myometrium. Paired samples of eight leiomyomas and adjacent myometrium were obtained and submitted to two-dimensional gel electrophoresis (2-DE) and mass spectrometry for protein identification and to Western blotting for 2-DE data validation. The comparison between the patterns revealed 24 significantly upregulated (p < 0.05) protein spots, 12 of which were found to be associated with the metabolic processes of the leiomyoma and not with the normal myometrium. The overexpression of seven proteins involved in the metabolic processes of the leiomyoma was further validated by Western blotting and 2D Western blotting. Four of these proteins have never been associated with the leiomyoma before. The 2-DE approach coupled with mass spectrometry, which is among the methods of choice for comparative proteomic studies, identified a number of proteins overexpressed in the leiomyoma and involved in several biological processes, including metabolic processes. A better understanding of the mechanism underlying the overexpression of these proteins may be important for therapeutic purposes.

Identification of proteins with different abundance associated with cell migration and proliferation in leiomyoma interstitial fluid by proteomics

“…This further confirms that glycolytic enzymes can be secreted by cells, attest metabolic dysregulation in leiomyomas (12) and perform non-enzymatic activities such as structural or regulatory functions at their extracellular localization (28).…”

“…In addition, the abundance of the proteins identified in our previous study (DES, TAGLN, SERPINA1, LMNA, ACTN1, PRDX2 and HSPA1A) is in line with the results of the present study on the leiomyoma IF. Furthermore, the levels of CRABP2, CALM1, LDHB, SERPINA1 and DES were confirmed to be upregulated, while those of TAGLN, LMNA and ACTN1 were downregulated, both in the proteome (12,13) and in the IF. Seven of these proteins (LUM, PRDX1, CRABP2, CALR, ANXA2, ALDOA and PGAM1) were validated by western blotting, thus confirming the 2-DE data.…”

“…Our previous proteomic studies on leiomyoma tissue identified several dysregulated proteins involved in metabolic processes, oxidative stress and cell migration (12,13). In a previous study, seven proteins with changes in abundance were identified in the IF of leiomyomas (14).…”

“…Spot quantification was performed as previously described (12). 2-DE image analysis was performed using the Proteomweaver 4 software.…”

Dysregulated chaperones associated with cell proliferation and negative apoptosis regulation in the uterine leiomyoma

“…Several external factors, including proteins, can induce alterations in the mechanotransduction signal from ECM via the transmembrane (8) receptor to the interior of the cell, leading to cell migration (9), metabolic dysregulation (10), proliferation and growth.…”