PAX2 Regulates PR-B Through AKT/mTOR Pathway In Endometrioid Carcinoma And Affects The Sensitivity Of Progesterone Therapy

Background:Progesterone therapy is often used as a conservative treatment for highly differentiated endometrioid adenocarcinoma in order to preserve the patient's reproductive function or avoid high-risk surgery.However,progesterone resistance has become a major clinical problem in conservative treatmentObjective:In order to confirm that PAX2 can affect the progesterone effect of endometrioid adenocarcinoma by regulating PR-B,and to further explore the mechanism of progesterone resistance of endometrioid adenocarcinoma,it provides a new molecular target for improving the drug efficacy of early endometrioid adenocarcinoma and preserving the reproductive endocrine function of patients.Methods:Part ?:1.qRT-PCR and Western blot was used to detect the original expression of PAX2 in 5 endometrioid cancer cells.2.After overexpressing/interfering with PAX2,progesterone was used to detect the ability of tumor proliferation,invasion and migration(CCK8,plate cloning,Transwell,scratch test)Part ?:1.After overexpressing/interfering with PAX2,progesterone was used to detect the expression of AKT/mTOR pathway protein by Western blot.2.COIP identified the interaction between PAX2 and PR-B.3.After overexpression of PAX2,progesterone was used to search for differential genes of downstream of PR-B by CHIP sequencingResults:1.The results of qRT-PCR and Western blot showed that the expression of PAX2 was relatively low in ISH,KLE and HEC cell lines and relatively high in RL and JEC cell lines2.The results of CCK8,plate cloning,Transwell and scratch assay showed that PAX2 overexpression combined with P4 treatment could effectively inhibit the proliferation,invasion and migration of ISH and KLE cell lines3.Western blot results showed that both overexpression of PAX2 and P4 treatment could inhibit AKT/mTOR pathway,but overexpression of PAX2 combined with P4 treatment had a more obvious inhibitory effect on AKT/mTOR pathway in ISH and KLE cell lines.Overexpression of PAX2 could promote the expression of PR-B in these two cell lines,but after the action of AKT/mTOR pathway promoter(IGF),the expression of PR-B in the overexpression of PAX2 was inhibited in ISH cell line4.The results of exogenous co-immunoprecipitation showed that PAX2 could bind to progesterone receptor PR-B in ISH cell line regardless of the presence of progesterone5.Chip-seq results showed that PAX2 could reduce the expression of ERBB2,the downstream gene of PR-B,in ISH cell line,and GEO data showed that the expression of ERBB2 in EEC was higher than that in normal tissues adjacent to cancer.Conclusion:1.PAX2 can indirectly promote the expression of PR-B by inhibiting AKT/mTOR pathway,thus enhancing the progesterone effect of EEC2.PAX2 can also enhance the sensitivity of EEC to progesterone by interacting with PR-B and regulating the expression of downstream genes of PR-B.