The Mechanism Of KDM1A Inhibitor Augments Efficacy Of Rapamycin For The Treatment Of Endometrial Cancer

Objective:Rapamycin is a classic inhibitor of mTOR,witch plays a important role in cell energy metabolism.KDM1A is type of transmethylase witch can regulate many biological processes,such as protein synthesis and DNA repair.KDM1A and mTOR were both found to be promote factors of tumorigenesis.The purpose of this study is to verify the role of KDM1A in endometrial cancer.Then explore the synergistic anti-cancer effect of KDM1A inhibitor and rapamycin.The possible mechanism of the synergistic anti-cancer effect is further explored.Method: Firstly,the KDM1A expression of endometrial cancer was analysed through ULCAN database.Then we used endometrial cancer lines,HEC1 A and RL95-2,and primary endometrial cancer cells from patients for cell experiments.KDM1A knockdown cell models was established by lentivirus infection,which content KDM1A sh RNA.We verified the role of KDM1A in endometrial cancer and the synergistic anticancer effect of KDM1A inhibitor and rapamycin the through proliferation,stemness and migration assay.Then we processed tumor 3D culture model and Cell line-derived xenograft model to further verified the conclusion.We collected all the tumor tissues for Immunohistochemistry(IHC),which can show the Ki67 and p-Akt expression level of tumor.The relation between KDM1A and PI3K/Akt/mTOR pathway was studied by TIMER2.0 database analysis.Finally we study the possible mechanism of the synergistic anti-cancer effect of KDM1A inhibitor and rapamycin through RNAseq analysis,real time PCR,chromatin immunoprecipitation assay(Ch IP)and Western Blot.Results: Bioinformatics analysis showed that KDM1A is highly expressed in endometrial cancer.Cell experiments showed KDM1A knockdown can inhibit proliferation,stemness and migration ability of cancer cell.And KDM1A inhibitor augments the efficacy of rapamycin for endometrial cancer treatment.The results of xenograft model experiment were in line with cell experiments.IHC assay showed KDM1A inhibitor and rapamycin treatment down regulate Ki67 and p-Akt expression of tumor.TIMER 2.0 database analysis showed KDM1A is positively relate to PI3K/Akt/mTOR pathway.RNAseq data showed KDM1A inhibitor and rapamycin can synergistically inhibit PI3K/Akt/mTOR pathway and down regulate lots of another biological processes like cyclin proteins,DNA repair and replication.Real time PCR,Ch IP and Western Blot were processed to verify the conclusion.Conclusions:(1)KDM1A is highly expressed in endometrial cancer.(2)KDM1A knockdown can inhibit proliferation,stemness,migration ability of cancer cell and augment the efficacy of rapamycin for endometrial cancer treatment.(3)KDM1A inhibitor and rapamycin have synergistic anti-cancer effect.(4)The possible mechanism of the synergistic anti-cancer effect of KDM1A inhibitor and rapamycin is that they can both inhibit PI3K/Akt/mTOR pathway.Besides that,down regulation of cyclin proteins,DNA repair and replication relate genes may play a role in this mechanism.