Eman Ibrahim Anwar^{1, 2}, Passant Mohamed Mohie², Suzan Elsharkawy³, Yasmine Amr Issa⁴, Lobna MMA Abd El Mottelib⁵, Rashed W Alweshah⁶, Ezzeldin Ahmed Hussein⁷ , Samar R Saleh^{8, 9}, Eman Helmy Thabet^{10, 11}

Abstract

Purpose: Endometriosis is a medical condition characterized by an endometrial-like epithelium and stroma outside the endometrium. It is generally managed with surgical intervention and hormonal therapies. This study sought to compare the relative potency of metformin and quercetin to cisplatin and their effects on proliferative and migratory characteristics of in vitro-cultured endometrial stromal cells derived from tissue samples of patients with endometriosis. Methods: Isolated endometrial mesenchymal stromal cells (EndMSCs) were obtained from forty patients with endometriosis at Shatby-Alexandria University Hospital, and expanded in vitro. The cells were validated using the cluster of differentiation marker (CD10) and estrogen receptor as positive markers and desmin as a negative marker. The MTT assay was conducted to assess the cytotoxicity of the three-drug formulations at different molar concentrations on EndMSCs, and their IC50 was determined. The following concentrations are used for Cisplatin (1 - 25 μM), Quercetin (1 -160 μM, and Metformin (1 - 40 mM). The markers of oxidative stress and Ki-67 expression were measured in the EndMSCs’ culture media, as well as the cell migratory potential. Extracellular signal-regulated kinase (ERK) and Phospho-ERK, a central dysregulated pathway in endometriosis, were assessed in EndMSCs using Western blot assay. Results: The calculated IC50 of cisplatin, quercetin, and metformin were 12.57 ± 1.635 μM, 261.2 ± 7.850 μM, and 92.01 × 10^3 ± 21.1 μM, respectively. Both metformin and quercetin-treated EndMSCs exhibited significantly reduced Ki-67 expression (p < 0.05 and p < 0.01), less cell migration profiles (p < 0.0001), decreased expression of pERK1/2 (p < 0.0001 and p < 0.001), with lower cell cytotoxicity (IC50) (p < 0.0001) and oxidative stress (p < 0.0001) compared to cisplatin. To the best of our knowledge, no studies have compared the in vitro effects of metformin, quercetin, and cisplatin on the proliferation and migration of EndMSCs. Conclusion: The findings of this study indicate that both drugs are promising agents for treating endometriosis. However, quercetin demonstrated higher potency in anti-proliferative and antioxidant effects, while metformin showed an obvious modulation of pERK1/2 to ERK ratio.