Effects of Vascular Endothelial Growth Factor(VEGF)Regulating Protein Kinase C Expression on the Invasiveness of Stromal Cells in Endometriosis
Objective To investigate the effects of vascular endothelial growth factor(VEGF)regulating protein kinase C(PKC)expression on the invasiveness of stromal cells in endometriosis(EMs).Methods Twenty patients with ovarian(EMs)and EMs confirmed by intraoperative and postoperative pathology were collected.The eutopic and ectopic endometrial tissues were collected and divided into EMs eutopic group and EMs ectopic group.The eutopic and ectopic endometrial tissues of 20 patients operated for ovarian endometriosis and pathologically confirmed as EMs after postoperative surgery were collected and divided into EMs.20 patients operated for other benign ovarian tumors in the same period were collected as the control group.Immunofluorescence staining was used to detect the expression and localization of VEGF and PKC proteins in the stromal cells of the EMs group.Western blot was used to detect the expression of VEGF and PKC protein in the control group and EMs group.EMs stromal cells were transfected by VEGF-siRNA gene silencing method,and the changes of VEGF and PKC protein expression after intervention were detected by Western blot.Transwell method was used to detect the invasiveness of cells after intervention.Results Compared with the control group,the expression of VEGF and PKC proteins in the EMs eutopic and ectopic groups increased,and the expression levels in the ectopic endometrium group were higher than those in the eutopic endometrium group(P all＜0.01).The results of cell immunofluorescence staining showed that green fluorescence was produced in both VEGF protein and PKC protein in the endometrial stromal cells of EMs.VEGF protein was expressed in both the cytoplasm and nucleus of the endometrial stromal cells,while PKC protein was mainly expressed in the cytoplasm of the endometrial stromal cells.Western blot results showed that compared with the control group,the expressions of VEGF and PKC proteins in the EMs group were increased(P all＜0.001).The results showed that the siRNA transfection efficiency was the highest after 72 h of culture.Western blot results showed that compared with siR-NC group,the expression of VEGF and PKC proteins in EMs interstitial cells was decreased after VEGF-siRNA intervention(P all＜0.001).Transwell results showed that compared with siR-NC group,after VEGF-siRNA intervention,the cell invasiveness of EMs interstitial cells was decreased(P all＜0.001).Conclusion VEGF-mediated vascular endothelial migration might be related to the expression of PKC protein,which induces the occurrence and development of EMs by changing the cell invasion activity.
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