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Abstract The purpose from this study was to highlight the role of stem cells in uterine fibroid growth. Fibroids represent the most common benign tumour in women and cause many gynecological disorders as bleeding, pain, infertility and abortion. They are a leading cause of hysterectomies in many countries and this causes an economic burden. We have conducted this study from june 2016 till September 2018 in collaboration with muenster university in Germany. It was performed on 18 cases who underwent hysterectomy or myomectomy either laparoscopically or abdominally. The side population cells from human leiomyoma exhibit the key features of tumor-initiating cells, opening up new possibilities of understanding the origin and developing new non surgical approaches for leiomyomas. A thorough characterization of primary leiomyoma and myometrial cells from uterine leiomyoma and myometrium adjacent tissues was conducted in this study. We have done cell culture procedures for evaluating the expression of stem cell markers by flowcytometry and RT-PCR. In the current study, we have investigated the potential dysregulation of SOX2, OCT4, NANOG, KLF4 and NOTCH1, COX2 by analyzing their expression by qPCR (Taqman) and α SMA, VM and MSI-1 (SYBR-green) and when comparing the expression of the studied primers on endometrium, myometrium, fibroid and endometriosis tissues respectively, significant statistical difference was only found in the expression of NOTCH1, NANOG, SOX2 and COX2. In our study, when measuring NANOG and SOX2 primer expression, they are less expressed in endometriosis than fibroid because the NANOG delta CT of endometriosis was significantly higher than those of myometrium and fibroid and the endometriosis’ delta CT in SOX2 was higher than the fibroid’s by about 4 and nearly equal to that of the endometrium and myometrium. Regarding NOTCH1 marker expression, it is expressed more in fibroid than myometrium, but it was comparable to that of the endometrium. So, gamma-secretase inhibitors are used in our study to inhibit this stemness-related molecular pathway. In our study, no significant statistical difference was found in the expression of Vementin, α SMA or MSI-1 in myometrium and fibroid. The impact of targeting the Notch signaling pathway with gamma-secretase inhibitors by Annexin V apoptosis assays, and MTT cell viability assays have been done. |