Fig. 5: Adipocytes mediated EV secretion increased EC cells proliferation/migration and EC progression. | Oncogene

Fig. 5: Adipocytes mediated EV secretion increased EC cells proliferation/migration and EC progression.

From: Obesity-induced extracellular vesicles proteins drive the endometrial cancer pathogenesis: therapeutic potential of HO-3867 and Metformin

Fig. 5

A Adipose-derived mesenchymal stem cells underwent adipocyte differentiation initiated 72 h post-confluency on Day 3, 5, and 7, followed by maintenance in adipocyte medium until day 15. Microscopic images (10x and 40x) at different time points show increasing lipid accumulation. B To confirm adipocyte formation, cells were cultured in a 12-well plate, fixed, and stained with 0.2% oil red O in 2-propanol for 10 min at room temperature. Adipogenesis was measured at various time points. C Adipocyte-mediated extracellular vesicle (EV) secretion and size confirmation by ISF. D Adipocyte-derived EV (ADEV) were co-cultured with IK cells for 48 h, and the expression of candidate proteins TMEM205, FAS, and STAT5 was analyzed by ELISA (n = 4, p < 0.01 or p < 0.005). E, F Cell proliferation assessed by 5-ethynyl 2´-deoxyuridine (EdU) incorporation using flow cytometry in IK cells co-cultured with ADEV, demonstrating an increased percentage of EdU+ve cells compared to IK control cells (untreated ADEV) (n = 3, p < 0.05 or 0.01). G Ishikawa (IK) cells (2 ×10^6 cells) were injected subcutaneously into the right flank of athymic nude mice. One week later, the mice were randomly divided into three groups: Cancer Control (IK cells only, n = 6), EV25µg group (IK cells + EV, n = 7), and EV100µG group (IK cells + EV, n = 7). Adipocyte-derived EV (25 µg and 100 µg per 100 µL) were injected subcutaneously near the tumors in the EV groups, while physiological saline was injected into the Cancer Control group. Injections were administered twice a week for 4 weeks. H, I Tumor growth was monitored weekly, and tumor size was measured using a linear digital caliper. Tumor volume was calculated and represented in the graph. J At the end of the ADEV treatment, the tumor-bearing mice were sacrificed. Tumor tissues were collected and subjected to ELISA to measure the expression of candidate proteins (TMEM205, STAT5, FAS, and Rab27a) in both the control and AEV-treated groups (n = 6; p < 0.001).

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