TGF-β Induced Epithelial-to-mesenchymal Transition Contributes To The Downregulation Of HSD17B2 Expression In Endometriosis Cell Model

Endometriosis is a benign disease induced by estrogen,characterized by ectopic growth of endometrioid tissue.Hydroxysteroid 17-beta dehydrogenase 2(HSD17B2)is a metabolic enzyme secreted by epithelial cells that converts estradiol into estrone,which is crucial for maintaining stable hormone levels in the body.Due to the special disease microenvironment of endometriosis,the expression of HSD17B2 in the lesions is reduced,mainly due to the lack of progesterone receptor(PR)due to abnormalities in the interstitial cells of the lesions,which prevents epithelial cells from secreting HSD17B2 normally through paracrine inducible factors.However,it is unclear whether there are other mechanisms for HSD17B2 in endometriosis.In this article,we use the eutopic endometrial epithelial cell 12 Z from female patients with peritoneal Endometriosis as the basic cell of this study,and use recombinant human transcription growth factor β(rhTGF-β)It induced epithelial to mesenchymal transition(EMT)of 12 Z cells.Through morphological observation of living cells,it was found that Morphogenesis of treated cells changed significantly compared with the control group,from tightly arranged stable epithelial cells to loosely disordered interstitial cells.Through real-time quantitative PCR,protein blotting and other methods to detect E-cadherin,N-cadherin and Vimentin,we can see the changes in the depth of protein blotting.According to the results of CT values and using Graph Pad Prism 9.0 software to plot the histogram,we found that the expression of E-cadherin on the surface of epithelial cells was significantly reduced,and the expression of N-cadherin,Vimentin and other proteins on the surface of interstitial cells was significantly increased.These markers are the key markers for EMT.Fluorescence intensity display and cell morphology observation using immunofluorescence technology revealed TGF-β In the treatment group,the fluorescence intensity of epithelial markers decreased,while the fluorescence intensity of interstitial markers increased,accompanied by changes in cell morphology.The results showed that EMT existed in Endometriosis cells.The expression of PR was further detected by protein blotting,and the histogram was drawn with Graph Pad Prism 9.0 according to the image J gray-scale scanning results.The results showed that compared with the control group,the expression of PR was reduced.Previous studies showed that in the lesions of Endometriosis,TGF-β The induced EMT is negatively correlated with the expression of PR,which is also a marker for distinguishing between normal uterus and endometriotic lesions.Therefore,we used rhTGF-β The cell model of Endometriosis was successfully constructed by inducing 12 Z cells.Subsequently,Western blotting was performed on the expression of HSD17B2 in the cell model,and the results showed that compared with the control group,TGF-β The expression of HSD17B2 decreased in the treatment group cells.This indicates that the expression of HSD17B2 in Endometriosis cell model will be reduced after EMT.After using si RNA to knock down EMT to activate the transcription factor snail,mesenchymal epithelial transformation(MET)occurred,and protein blotting results showed a callback in the expression of HSD17B2.After knocking down the EMT transcription activator slug,there was no change in the expression of HSD17B2.This result indicates TGF-β is present in the endometriotic cell model activating snail transcription factor to initiate EMT reduces the expression of HSD17B2.In summary,we demonstrated tha this article demonstrates TGF-β could induce a cell model of endometriosis EMT in endometriosis histiocyte,which reduced the expression of HSD17B2.