ABSTRACT
Background: Endometriosis is a chronic inflammatory condition characterized by the presence of endometrial tissue outside the uterus, which can cause pelvic pain, infertility, and other symptoms. The disease may manifest as superficial peritoneal or deep-infiltrating endometriosis or as ovarian endometriomas. Although the mechanisms associated with the regulation and production of inflammatory mediators in endometriosis have been widely investigated, the precise mechanism responsible for inflammation-induced pain remains unclear, and the findings related to the cytokine expression profile and the location of cytokines in cells are contradictory. The intensity of pain experienced by endometriosis patients is not proportional to the degree and severity of their disease. Pain has a significant impact on women suffering from endometriosis. Methods: The following inclusion criteria to the study were: presence of endometriomas vs teratomas, negative pregnancy test result, no prior obstetric and infertility treatment, and good health condition with no diseases or coagulation disorders. Blood samples were collected from all patients. The serum levels of chemokines were determined by ELISA. The Nottingham Health Profile (NHP) questionnaire was made. Results: The median serum levels of chemokines: Monocyte Chemoattractant Protein 1 (MCP-1) and Monocyte Chemoattractant Protein 3 (MCP-3) were statistically higher in the endometriomas group compared to the other two groups. In the NHP questionnaire the comparison of the subjective health dimensions in individual groups showed that the patients in the endometriomas group experienced a significantly higher intensity of "PAIN" compared to other groups. Correlation analysis between NHP dimensions and serum chemokine levels: spearman's rank correlation analysis indicated a statistically significant relationship between the "VITAL ENERGY" dimension and the level of MCP-2 (r = -0.295;p = 0.022), MCP-3 (r = 0.254;p = 0.050), and RANTES (r = -0.353;p = 0.006);between the "EMOTIONS" dimension and the level of MCP-3 (r = 0.262;p = 0.043);and between the "INCONVENIENCE IN DAILY LIFE" dimension and the level of Eotaxin-1 (r = -0.283;p = 0.028) and CCL13 (r = -0.287;p = 0.026). Conclusions: The chemokines serum levels (i.e., MCP-1 and MCP-3) and intensity of "PAIN" were statistically higher in the endometriomas compared to the teratomas group of women. Therefore, understanding their role in endometriosis-related pain could help in the development of novel, multidisciplinary treatments.
ABSTRACT
Proteasome inhibitors are among the most potent classes of drugs in multiple myeloma treatment. One of the main challenges in myeloma therapy is acquired resistance to drugs. Several theories have been proposed to describe the mechanisms responsible for resistance to the most commonly used proteasome inhibitors bortezomib and carfilzomib. This study aimed to describe functional differences between sensitive myeloma cells (MM1S WT) and their daughter cell lines resistant to either bortezomib (MM1S/R BTZ) or carfilzomib (MM1S/R CFZ), as well as between both resistant cell lines. Bortezomib- and carfilzomib-resistant cell lines were successfully generated by continuous exposure to the drugs. When exposed to different drugs than during the resistance generation period, MM1S/R BTZ cells showed cross-resistance to carfilzomib, whereas MM1S/R CFZ cells were similarly sensitive to bortezomib as MM1S WT cells. Following proteomic profiling, unsupervised principal component analysis revealed that the MM1S/R BTZ and MM1S/R CFZ cell lines differed significantly from the MM1S WT cell line and from each other. Canonical pathway analysis showed similar pathways enriched in both comparisons - MM1S WT vs. MM1S/R CFZ and MM1S WT vs. MM1S/R BTZ. However, important differences were present in the statistical significance of particular pathways. Key alterations included the ubiquitin-proteasome system, metabolic pathways responsible for redox homeostasis and the unfolded protein response. In functional studies, both drugs continued to reduce chymotrypsin-like proteasome activity in resistant cells. However, the baseline activity of all three catalytic domains of the proteasome was higher in the resistant cells. Differences in generation of reactive oxygen species were identified in MM1S/R BTZ (decreased) and MM1S/CFZ cells (increased) in comparison to MM1S WT cells. Both baseline and drug-induced activity of the unfolded protein response were higher in resistant cells than in MM1S WT cells and included all three arms of this pathway: IRE1α/XBP1s, ATF6 and EIF2α/ATF4 (downstream effectors of PERK). In conclusion, contrary to some previous reports, resistant MM1S cells show upregulation of unfolded protein response activity, reflecting the heterogeneity of multiple myeloma and prompting further studies on the role of this pathway in resistance to proteasome inhibitors.