| BackgroundEndometriosis(EMs)was defined as endometrial tissues(gland and stroma)appear outside the uterus.Endometriosis is a common benign gynecological disease in women,but it can exhibit malignant biological behaviors which are similar to tumors.In recent years,domestic and foreign scholars have proposed a variety of theories including the theory of menstrual blood reflux,the theory of lymphatic and venous dissemination,the theory of iatrogenic implantation,the theory of body cavity metaplasia and the theory of intima determinism with respect to the development of endometriosis,but its specific pathogenesis still remains unclear and needs further researches.Collagen triple helix repeat containing-1(CTHRC1),a 28 k D secreted glycoprotein,was originally found in differentially expressed genes in injured rat balloon arteries.It is mainly distributed in the intercellular space,blood vessels,gastrointestinal tract and uterine smooth muscle,which can promote the repair process of injured tissues by inhibiting collagen synthesis and inducing cell migration.Many studies in recent years have shown that CTHRC1 is highly expressed in a variety of tumor cells,including breast cancer,liver cancer,non-small cell lung cancer,pancreatic cancer,and melanoma.Its abnormally high expression promotes tumor development by inducing tumor cells migration and invasion,promoting angiogenesis and tumor metastasis.Although studies have demonstrated that CTHRC1 plays an important role in the pathogenesis of manytumors,it remains unclear whether CTHRC1 is involved in the development of endometriosis which can exhibit malignant biological behaviors similar to tumors.The purpose of this study was to examine and analyze the expression of CTHRC1 in patients with and without endometriosis and study the effects of CTHRC1 on the migrative,invasive and proliferative abilities of ectopic endometrial stromal cells.Furthermore,to explore the underlying molecular mechanism of CTHRC1 in endometriosis and provide a new potential molecular target for clinical treatment of endometriosis.ObjectiveTo study the role of CTHRC1 in the pathogenesis of endometriosis.To elucidate the effect of CTHRC1 on the migration,invasion and proliferation of ectopic endometrial stromal cells,and to explore its underlying molecular mechanism.Materials and methodsTotal 36 tissue specimens were collected from endometrium of non-endometriosis and endometriosis patients,and ovarian ectopic cyst specimens of endometriosis patients.The m RNA and protein levels of CTHRC1 in the three groups were detected by RT-q PCR and Western Blot.Preoperative serum samples from non-endometriosis and endometriosis patients were collected,and the concentration of secreted CTHRC1 protein in serum samples was detected by ELISA.4 cases of ovarian ectopic cysts from endometriosis patients were collected and cultured.Primary ectopic endometrial stromal cells(EESCs)were isolated and immunofluorescence was used to detect cell purity.EESCs were transiently transfected with small interfering RNA(si RNA-CTHRC1)to inhibit CTHRC1 expression.Western Blot and RT-q PCR were used to detect the transfection efficiency.The si RNA-CTHRC1 treatment group and the negative control treatment group were processed to detect the change of cell migrative and invasive abilities by wound scratch assay and Transwell assay.The cell proliferative ability was detected by CCK8 assay,and the proportion of cell cycle was detected by flow cytometric assay.Western Blot were used to explore the changes in activity of theWnt/β-catenin pathway.Results1.The expression level of CTHRC1 in ovarian ectopic cyst specimens ofendometriosis patients is significantly higher than that in endometrium ofendometriosis and non-endometriosis patients.The expression level of secretedCTHRC1 in serum samples of patients with endometriosis is significantly higherthan that of non-endometriosis patients.2.Ectopic endometrial stromal cells were successfully isolated and cultured,whichhad a long spindle shape under microscope and were stable in status and can bepassed on.3.After inhibiting the expression of CTHRC1,the migration,invasion andproliferation of EESCs were significantly decreased.The proportion of cells in G1phase was significantly increased,and the proportion of S phase was significantlydecreased.4.The activity of Wnt/β-catenin signaling pathway was significantly decreased inEESCs after inhibiting the expression of CTHRC1.Conclusion1.Abnormal expression of CTHRC1 might be associated with the development ofendometriosis.2.CTHRC1 may affect the migration,invasion and proliferation of EESCs throughWnt/β-catenin signaling pathway. |
