The Effects Of IL-17A On Proliferation Of Human Endometrial Cells In Vitro

Objective: In this study,the Interleukin-17 A in different concentrations was applied to the endometrial cells cultured in vitro from normal endometrial tissues and EM patients’ eutopic endometrial tissues to observe the effect of IL-17 A on proliferation of human endometrial cells in the simulative microenvironment of endometriosis.Methods:1 The human endometrial epithelial cells(EECs)and endometrial stromal cells(ESCs)were isolated and cultured in vitro by collagenase I digestion.The morphology and growth status of the cells was observed using the inverted phase contrast microscope.EECs and ESCs were identified by immunocytochemistry.2 The IL-17 A receptors on human endometrial cells were detected by immunochemistry.3 The IL-17 A in different concentrations was applied to EECs and ESCs for 24 h to observe the effect on proliferation of the cells.The MTT experiments were performed to select the optimal concentration for subsequent experiments.4 The mRNA of PCNA gene in EM patients’ eutopic endometrial cells treated by IL-17 A in the concentrations of 0μg/ml and 250μg/ml was detected by real-time PCR and compared by statistical analysis.5 The expression of β-catenin in EM patients’ eutopic endometrial cells treated by IL-17 A in the concentrations of 0μg/ml and 250μg/ml was detected by western-blot and compared by statistical analysis.Results:1.There was no significant difference in the morphology of normal endometrial cells and EM patients’ eutopic endometrial cells.The adherentEECs grew in the form of onion or crumb.The results of immunocytochemistry showed that the Cytokeratin in cytoplasm was stained to claybank,which was positive.The non-staining nucleus was stained purple by hematoxylin.And the stain of Vimentin was negative.Most of the ESCs dispersed to single cells.The adherent cells in the form of polygon were arranged in bundles.The results of immunocytochemistry showed that Vimentin in cytoplasm was stained to claybank.The non-staining nucleus was stained purple by hematoxylin.The stain of Cytokeratin was negative.2.The results of immunocytochemistry of EECs and ESCs from normal endometrial tissue and EM’s eutopic endometrial tissue showed that the cytoplasm was claybank,which indicated positive expression.The nucleus was nonstaining,which was negative.3.There was no significant difference between each treatment and control groups in MTT by treating human normal endometrial cells for 24 h using IL-17 A in different concentrations,P>0.05.4.The endometrial cells from EM patients were treated by IL-17 A in different concentrations for 24 h.The OD in each treatment groups was compared with that in control group.The OD in each treatment group was significantly higher than that in control group except for the group of IL-17 A in 50μg/ml(P<0.05).The OD differences between treatment groups were not significant.5.The results of RT-PCR showed that the level of PCNA mRNA in EECs and ESCs in the treatment group was significant higher than that in control group(P<0.05).6.The western-blot results showed that the expression of β-catenin in EECs and ESCs in treatment groups were significantly higher than that in control group(P<0.05).Conclusions:1.There is no visible difference in the morphology of normal endometrial cells and EM patients’ eutopic endometrial cells.2.IL-17 A has no effect on normal human endometrial cell proliferation.3.IL-17 A can promote the expression of PCNA mRNA in the epithelial cells and stromal cells from EM patients’ eutopic endometrial tissues.And the expression of PCNA mRNA in stromal cells is higher than that in epithelial cells.4.IL-17 A can promote the expression of β-catenin in the epithelial cells and stromal cells in EM patients’ eutopic endometrial tissues.And the expression of β-catenin in stromal cells is higher than that in epithelial cells.5.IL-17 A can promote the proliferation of EM patients’ eutopic endometrial cells and may be involved in the development of endometriosis..