Researches About LncRNA AFAP1-AS1 Promotes The Development Of Endometriosis Through The Wnt5a-RhoA Pathway

Endometriosis is a gynecological disease that seriously endangers women's health.However,the aetiology and pathogenesis of endometriosis remain incompletely understood.Many studies have revealed that IncRNAs(long non-coding RNA)play important roles in the occurrence and development of endometriosis.Previously,we found that the expression of lncRNA AFAP1-AS1 was higher in ectopic endometrial tissues than that in normal endometrial tissues.Moreover,the down-regulation of AFAP1-AS 1 had an inhibitory effect on the occurrence and development of endometriosis,but the mechanisms remain unknown.In this study,the function of AFAP1-AS1 in endometriosis was verified;and the expression of Wnt5a-RhoA signaling pathways related molecules in endometriosis was detected;and the effects of RhoA on biological function of eutopic endometrial stromal cells were explored;and the changes of gene and protein expression of Wnt5a-RhoA signaling pathways related molecules in endometriotic stromal cells after AFAP1-AS1 knockdown were observed;and effects of exogenous Wnt5a protein and RhoA inhibitor on the biological function of endometriotic stromal cells after AFAP1-AS1 Lknockdown were studied.Finally,effects of estrogen on the expression of AFAP1-AS1 and its downstream molecules were observed.The main results were as follows:(1)AFAP1-AS1 is highly expressed in ectopic tissues,which promotes the migration and invasion of endometriotic stromal cells.(2)The gene and protein expression of Wnt5a,RhoA,ROCK1 and ROCK2 in ectopic tissues were significantly higher than those in normal endometrium.The gene expression of Wnt5a and RhoA in ectopic tissues was significantly related to AFAP1-AS1.The activator and inhibitor of RhoA promoted and attenuated the proliferation,migration,and invasion abilities of eutopic endometrial stromal cells,respectively.(3)The gene and protein expression of Wnt5a,RhoA,ROCK1 and ROCK2 decreased after AFAP1-AS1 knockdown in endometriotic stromal cells.Wnt5a protein and RhoA inhibitor promoted and attenuated the proliferation,migration and invasion of endometriotic stromal cells after AFAP1-AS1 knockdown,respectively.However,the promoting effect of Wnt5a was offset by the downstream RhoA inhibitor when acting together.(4)Estrogen promoted the expression of AFAP1-AS1 and its downstream proteins in normal endometrial stromal cells.In summary,the expression of AFAP1-AS1 and Wnt5a-RhoA signaling pathways related molecules were both increased in ectopic tissues of endometriosis,and RhoA promotes the development of endometriosis.AFAP1-AS1 promotes the occurrence and development of endometriosis,and its molecular mechanism is related to the Wnt5a-RhoA signaling pathways.Estrogen can promote the expression of AFAP1-AS1 and its downstream proteins in normal endometrial stromal cells,suggesting that the regulation of AFAP1-AS1 in endometriosis is closely related to estrogen.