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

Abstract

Abstract. Uterine leiomyoma is the most common female reproductive tract benign tumor. Little is known about protein composition and changes in the leiomyoma interstitial fluid (IF). The present study focused on changes in protein abundance in the IF of leiomyoma. Leiomyoma IFs and adjacent myometrial IFs were obtained and analyzed by two-dimensional electrophoresis (2-DE) coupled with mass spectrometry and western blotting for 2-DE data validation. A total of 25 unique proteins were observed to change significantly (P<0.05). Of these proteins with different abundance, 22 had not been previously identified in leiomyoma IF. In silico analysis predicted that three of these proteins were secreted via classical mechanisms, while 22 were secreted via non-classical mechanisms. Ingenuity Pathway Analysis identified 17 proteins associated with cellular migration and proliferation. Among these, phosphoglycerate mutase 1 had not been previously associated with leiomyoma. The abundance of seven proteins was further validated by western blotting. A comparative proteomic approach identified a number of proteins associated with cellular migration and proliferation, with changes in abundance in IF likely to be involved in tumor development. Further studies will be required to investigate the role of these proteins in leiomyoma IF and their possible association with tumor development and growth.

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

“…In this study we found HSPA1A and HSPB1 to be downregulated in the leiomyoma if compared to the myometrium. In our previous study (8) we identified HSPA1A as less abundant in the leiomyoma interstitial fluid, this being in line with our present data. HSPB1 belongs to the group of small Hsps with anti-apoptotic and tumorigenic properties, and considered as important therapeutic targets, particularly in cancer pathology (25,26).…”

“…The expression of PDIA3, HSPA5, CALR, was confirmed to be upregulated, while the expression of HSPA1A and HSPB1 was confirmed to be downregulated. The abundance of VLC was confirmed to be upregulated in the leiomyoma interstitial fluid (8).…”

“…Trypsin digestion and MALDI analysis. Spots from 2-DE were digested with sequencing grade-modified trypsin (Promega Corporation, Madison, WI, USA) and analyzed by mass spectrometry (MS), as described by Ura et al (8).…”

“…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.…”

Interstitial Fluid in Gynecologic Tumors and Its Possible Application in the Clinical Practice

“…The presence of intracellular proteins is not a problem for biomarker identification, but may be a challenge for the identification of proteins actually present in the TIF. In gynecology, this method was used in ovarian cancer [71,72,73] and leiomyoma [74,75] for the exploration of the TIF by proteomics.…”