The Study Of The Mechanism Of Artemisinin Derivative DHA/ARTS Inducing Ferroptosis In Renal Carcinoma Cells 786-0

Objective: To explore the mechanism of artemisinin derivative DHA/ARTS inducing ferroptosis in renal cancer cell line 786-0.Methods: DHA/ARTS stimulate renal cancer cell line 786-0,CCK-8 was used to detect the effect of DHA/ARTS to kill 786-0 cell,the intracellular ROS level was detected,and ferroptosis inhibitors DFO/Fer-1 was used to observe its effect on DHA/ARTS inhibition,determine the death mode of DHA/ARTS-induced renal cancer cell death;using western blot and other experimental techniques to observe the changes in protein levels of NCOA4 and FTH1 after DHA/ARTS inducing;after knockdown NCOA4 or NLK with two different siRNAs,observe the change of DHA/ARTS effect on 786-0;construct and transfect pmCherry-EGFP-FTH1 plasmid and fluorescence coloration,observe the change of fluorescence color and brightness after DHA/ARTS effect,judge the autophagic degradation of FTH1;knockdown NCOA4 with different siRNAs,transfect the pmCherry-EGFP-FTH1 plasmid to detect the change of fluorescence color and brightness after the effect of DHA/ARTS,and explore the regulation of FTH1 degradation by NCOA4.Results: Through cell experiments,it was confirmed that DHA and ARTS can induce the increase of intracellular ROS levels,leading to the death of 786-0 cells,and this death mode can be inhibited by the ferroptosis inhibitors DFO/Fer-1;DHA and ARTS stimulate 786-0,NCOA4 decreases in protein level and is time-dependent;FTH1 protein expression also decreases,which is consistent with NCOA4;knockdown of NCOA4 or NLK can inhibit the killing effect of DHA and ARTS on 786-0 cells;overexpression of pmCherry-EGFP-FTH1 plasmid,after stimulating of DHA and ARTS,the yellow fluorescence decreases,the red fluorescence increases,and FTH1 autophagy degradation increases;after NCOA4 knockdown,the FTH1 autophagy degradation caused by DHA and ARTS is significantly reduced.Conclusion: Artemisinin derivatives DHA and ARTS can induce ferroptosis in renal cancer cell lines 786-0,and the ferroptosis is achieved through NCOA4-mediated autophagy degradation of FTH1.