| Endometriosis is a common,benign,estrogen-dependent chronic gynecological disease defined as the presence of functional endometrial tissue in the endometrium and other areas of the uterine body other than the muscularis.Endometriosis is characterized by pain associated with the menstrual cycle,such as dysmenorrhea,dyspareunia,abdominal pain,chronic pelvic pain,and/or infertility.Endometriosis mainly affects women of childbearing age,about 5%-10% of women of childbearing age are affected,and the prevalence rate is particularly high in infertile women,up to 50%.Postmenopausal endometriosis is relatively rare.In 1925,Sampson first reported an example of endometriosis malignancy and described its association with ovarian cancer.After a follow-up study,scientists found that endometriosis associated ovarian cancer generally includes ovarian clear cell cancer and ovarian endometrioid cancer,but the pathogenesis is still unclear.Previous literature review of this project shows that the expression of mTOR is elevated in endometriosis and ovarian clear cell carcinoma,but there are no relevant studies on normal endometrium and ovarian endometrioid carcinoma.Objective: This study aims to elucidate the changes and regulatory functions of mTOR complex mTORC1/2 in normal endometriosis,ovarian endometriosis cyst and endometriosis related ovarian cancer,and to preliminatively explore the clinical transformation value.Methods:1.According to the clinical data of Xiangya Hospital,Central South University,the expression of mTOR in endometriosis associated ovarian cancer in public database was analyzed by bioinformatics.2.To describe the changes of mTORC1/2 in normal endometrium,ovarian endometriosis and endometriosis related ovarian cancer,and to describe the relationship between the changes in expression and the stage and prognosis of endometriosis related ovarian cancer.(1)Pathological tissue and clinical data after surgical treatment were collected from Xiangya Hospital of Central South University to study the difference in prognosis between endometriosis related ovarian cancer and other ovarian cancers.(2)The expression of RAPTOR,p70s6 K,RICTOR and PKCa in normal endometriosis,ovarian endometriosis and endometriosis related ovarian cancer tissues was verified by immunohistochemistry.Western blotting was used to detect the expressions of RAPTOR,p70s6 K,RICTOR and PKCa in normal endometrial cell lines,ovarian clear cell cancer cells and ovarian endometrioid cancer cells.(3)To explore the correlation between immunohistochemical and endometriosis related ovarian cancer stage and prognosis.3.Proved the effect of mTORC1/2 on cell function in ovarian clear cell cancer cell lines and ovarian endometrioid cancer cell lines.(1)siRNA was used to knock out mTORC1/2 related regulatory proteins in ES-2 and TOV-112 D cell lines and verify the knockout effect(2)Verify the effect of mTORC1/2 on related cancer cells from cell proliferation rate and migration capacity.Results:1.By comparing the collected clinical data,it can be found that compared with ovarian serous cancer and ovarian mucinous cancer,endometriosis-associated ovarian cancer has an earlier stage of disease onset(P < 0.05).Further KM survival analysis shows that compared with ovarian serous cancer and ovarian mucinous cancer,the progression-free survival and overall survival of endometriosis associated ovarian cancer are significantly better(P < 0.05).Then R software was used to screen and compare differentially expressed genes from GEO database.In dataset GSE189553,it was found that compared with ovarian serous cancer,the expression of mTOR in endometriosis associated ovarian cancer was significantly increased(P < 0.05).Meanwhile,in another dataset GSE157153,the expression of mTOR in endometriosis-associated ovarian cancer was also significantly higher than that in paracancer endometriosis and endometriosis tissues(P < 0.05).2.Immunohistochemical detection of mTORC1 regulatory protein RAPTOR and its downstream p70s6 k and mTORC2 regulatory protein RICTOR and its downstream PKCa were performed using the collected tissue sections.It was found that the expression of these four proteins increased gradually in normal endometrium,ovarian endometriosis and endometriosis associated ovarian cancer(P < 0.05).WB experiment also confirmed that mTOR related regulatory protein was highly expressed in endometriosis associated ovarian cancer.At the same time,immunohistochemical tests showed that there was no significant difference in the expression of MTOR-related regulatory proteins between endometriosis tissues and their endometrium(P > 0.05).Immunohistochemical scores were further correlated with the disease stage,and it was found that the expression level of RAPTOR was positively correlated with the disease stage(P < 0.05),but the immunohistochemical scores of the four proteins were not significantly correlated with the prognosis of endometriosis-associated ovarian cancer.3.siRNA was used to successfully knock down RAPTOR and RICTOR in ovarian clear cell cancer cell line ES-2 and ovarian endometrioid cancer cell line TOV-112 D,and the knock-down effect was verified by WB.The cell proliferation rate was detected by CCK-8method.Compared with the control group,the cell proliferation rate in knockdown group was significantly decreased(P < 0.05).transwell was used to detect the migration and invasion ability of cells.Compared with the control group,the migration ability of cells in knockdown group was decreased(P < 0.05),and the invasion ability of TOV-112 D was significantly decreased(P < 0.05).Conclusion:1.Compared with other types of ovarian cancer,endometriosis associated ovarian cancer has an earlier onset stage and may have a better prognosis.2.mTOR-related regulatory proteins are highly expressed in endometriosis associated ovarian cancer and may be related to disease stage.3.mTOR-related regulatory proteins can promote the proliferation,migration and invasion of ovarian cancer cells associated with endometriosis.There are 24 figures,8 tables and 84 references... |
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