P265. Identification of kisspeptins and matrix metalloproteinases in endometrial cell culture from patients with endometriosis

Tatiana Kleimenova (RU), Anna Drobintseva (RU), Victoria Polyakova (RU), Valeriia Rodichkina (RU)

[Kleimenova] Federal State Budgetary Scientific Institution “The Research Institute of Obstetrics, Gynecology and Reproductology named after D.O.Ott” Сокращенное название: ФГБНУ «НИИ АГиР им. Д.О. Отта» FSBSI “The Research Institute of Obstetrics, Gynecology and , [Drobintseva ] Federal State Budgetary Scientific Institution “The Research Institute of Obstetrics, Gynecology and Reproductology named after D.O.Ott”, [Polyakova ] Federal State Budgetary Scientific Institution “The Research Institute of Obstetrics, Gynecology and Reproductology named after D.O.Ott”, [Rodichkina] Federal State Budgetary Scientific Institution “The Research Institute of Obstetrics, Gynecology and Reproductology named after D.O.Ott”

Objective: Endometriosis is a common condition where tissue that behaves like the lining of the womb (endometrium) is found in other parts of the body. The endometrium consists of a single-layered prismatic epithelium, uterine glands, and a specialized, cell-rich connective tissue (stroma). Kisspeptin (KISS) and kisspeptin receptor (KISS1R) regulate the function of metalloproteinases 9 (MMP-9), which play an important role in the invasion processes. Despite this, KISS/KISS1R activity in endometriosis has not investigated yet. Methods: The cells culture was isolated by enzymatic digestion (collagenase II, Gibco, USA) from human endometrial samples. Incubated at 37Co in DMEM/F-12 supplemented with 10% FBS. Immunocytochemistry (ICC) was done using the incubation with a 1:100 dilution of anti-human KISS1 monoclonal antibody (Abcam) and 1:200 dilution of anti-human GPR54 polyclonal antibody (Abcam). The primary antibody was omitted for negative controls. ICC assays were performed from the first passage. Patients: Human endometrial tissues were obtained following written consent for the use of surgical specimens from patients who underwent abdominoscopy or needle biopsy under protocols. Endometrial tissues from 4 donors with normal menstrual cycles (mean age 31.1). Patients having stage II endometriosis genitalis externa (n=5) (mean age 32.7), according to the AFA. Isolation of endometrial cells was carried out for 21-22 day of a menstrual cycle at 6 patients and for 7-12 day in 2 cases. For statistical analysis, Mann–Whitney U test was performed using Statistica 7.0 software. Results: KISS1 expression was detected in the culture studied. In the control group (S=1,02-1,41) reaction was higher than in the endometriosis (S=0,92-0,57), but this results not statistics (p >0,05). The levels of KISS1R did not differ between the studied groups (S=0,95-2,1). MMP-9 exhibited weak expression in cell culture (S=1,13-1,58). Conclusions: Expressions of KISS1/KISS1R and MMP-9 were found. Endometrial cell invasion frequently shares invasive characteristics that are inherent to cancer. Our intention is that KISS1 expression was inversely associated with stage of endometriosis. So, synthetic KISS could be used to suppress MMP activity in endometrial cells. The obtained cell culture can be used as a model for studying of endometriosis.