Inhibition Of Bladder Cancer Cell Proliferation And Mechanism By Sesquiterpene Lactones Of Elephantopus Scaber L.

Background: Scabertopin(ST)and Deoxyelephantopin(DET)are the main active components of Elephantopus scaber L with significant anticancer activity.Objective: To evaluate the antitumor activity of sesquiterpene lactone compounds(ST and DET)in bladder cancer and its potential molecular mechanism in vitro.Methods: Firstly,cell proliferation was detected by CCK-8 after acting on bladder cancer cells(J82 and T24)and non-cancerous cells(SV-HUC-1)by ST and DET.Next,J82 was selected as a further study,and cellular activity was detected using LIVE/DEAD,changes in cell morphology and organelles were observed utilizing SEM and TEM,intracellular ROS levels were detected by ROS fluorescent probes,and the effects of cell function were analyzed by cell cycle,apoptosis,migration and invasion.The expression of proteins(Caspase-3,Bax,Bcl-2,FAK,p-FAK,MMP-9,GPX4,GSDMD,RIP1,p-RIP1,RIP3,p-RIP3,MLKL and p-MLKL)was detected by WB.Results: ST and DET significantly inhibited the proliferative activity of bladder cancer cells T24 and J82.shortened and denser pseudopods,induction of intracellular ROS accumulation,inhibition of cell migration and invasion,induction of cell arrest in S and G2/M phases,upregulation of cell necrosis,and downregulation of p-FAK and MMP-9 expression were observed after ST and DET treatment of J82 cells,while Caspase-3,Bax and Bcl-2 expressions were unchanged.In addition,ST and DETtreated cells showed features such as cytoplasmic vacuolization and organelle swelling,as well as significantly increased phosphorylation levels of necroptosis-related proteins RIP1,RIP3 and MLKL.Conclusion: ST and DET induced the accumulation of ROS levels in bladder cancer J82 cells and decreased the expression of p-FAK and MMP-9 to inhibit cell migration and invasion;ST and DET also mediated necrotic apoptosis through activation of RIP/RIP3/MLKL phosphorylation and inhibited cell proliferation and viability.