Role Of Epidermal Growth Factor In HAS2/HA Pathway In Endometriosis

Background:Characterized by the presence of endometrium outside the uterine cavity,endometriosis is a benign gynecological disease,it has certain malignant biological behaviors such as proliferation,adhesion,invasion and metastasis.The progression of endometriosis is often associated with the implantation,dissemination,invasion,and local cytokine production and secretion of pelvic endometrial cells.Hyaluronan acid(HA)is an important tissue component in the extracellular matrix and is mainly synthesized by hyaluronan synthase 2(HAS2).The accumulation of HA paly an important role in the study of the mechanism of endometrial carcinogenesis.It has been found to have a certain role and the increase in HA concentration in many other cancers is associated with poor progression and poor survival.Silencing HAS2 in breast cancer studies has been shown to reduce HA synthesis and inhibit breast cancer cells migration and invasion.Studies have found that certain growth factors or hormones can modulate HAS2 at the transcriptional level.For example,epidermal growth factor(EGF)in epidermal keratinocytes can activate HAS2 and increase the expression of cellular hyaluronan.The literature reports that the EGF system is associated with the periodic growth and shedding of human intima,and there is a certain correlation between the severity of the study of endometriosis.After binding to the EGF receptor,EGFR is phosphorylated and subsequently transmitted through a series of transcription factors.Considering that EGF/HAS2/HA pathway may be involved in the development of endometriosis,the specific molecular mechanisms involved are still unknown,so it is worth further investigation.This study investigated the role and importance of EGF,HAS2,and HA in the pathogenesis and development of endometriosis from the aspects of molecular level regulation and cell migration.Method:Fresh EuEM tissue were used to culture primary cells.EuESCs were isolated and immunofluorescence staining was performed to detect the purity of mesenchymal cells.RT-qPCR was used to detect the effect of time and concentration of EGF on the expression of HAS2 mRNA in EuESCs.The effect of EGF on the expression of HA in the extracellular matrix of EuESCs was detected by ELISA.The short-term HAS2 expression inhibition system was established with siHAS2.EGF/DMSO and siRNA(siCtrl/siHAS2)were applied to EuESCs,and transwell was used to detect the effect of EGF and siHAS2 on the migration of EuESCs.Result:1.EuESCs were successfully cultured and the purity of the mesenchymal cells was above 95%.2.EGF can increase the mRNA expression of HAS2 in EuESCs,and has time and concentration dependence.3.EGF can increase the expression of HA in the extracellular matrix of EuESCs.4.Transfected EuESCs with siHAS2,transfection efficiency was greater than 70%;EGF in siCtrl group increased HAS2 expression,while EGF in siHAS2 group could not increase HAS2 expression.5.EGF can promote the migration of EuESCs.Interfering with the expression of HAS2 can partially inhibit the migration of EuESCs.Conclusion:EGF can increase the expression of HAS2 in EuESCs in a time-and concentration-dependent manner,which in turn increases the amount of HA in the HAS2 synthesis product in the extracellular matrix.EGF can promote the migration of EuESCs.Interfering with the expression of HAS2 can partially inhibit the migration of EuESCs.