Depletion Of Ferritin Light Chain(FTL)in Glioblastoma Induce Apoptosis And Autophagy And Its Underlying Mechanism

Objective: Ferritin light chain(FTL)is one of the two important subunits that make up ferritin.Studies have shown that it is highly expressed in glioblastoma cells and tissues,and its expression is negatively related to the clinical survival rate of patients.Important Physiopathological Mechanisms into Glioblasts Glioblastomas are the most common malignant tumors of the central nervous system.At present,there are no studies on the apoptotic autophagy of ferritin light chains.The purpose of this study was to investigate the relationship between FTL on apoptosis and autophagy in human glioblastoma,and to study the effects of down-regulation of FTL on human glioblastoma apoptosis and autophagy at the cellular level and its possible mechanism.Method: si RNA transfection technology was used to silence FTL expression in U251 and U87;real-time quantitative PCR was used to detect FTL and p53 m RNA expression in FTL silencing and control groups.Flow cytometry was used to detect changes in apoptosis,mitochondrial membrane potential,reactive oxygen species,and cell cycle in the FTL-silenced group and the control group;Western blot was used to detect caspase3,clevered-caspase3,Bax,Bcl-Expression of 2,P53,P62 and LC3 I /II;TUNEL test to detect apoptosis in FTL silencing group and control group;immunofluorescence test to detect clevered-caspase3,P53,P62 and LC3 I / in silencing group and control group Expression of II.Results:In glioblastoma cells U251 and U87,after si RNA reduced ferritin light chain,real-time quantitative PCR showed that FTL was successfully silenced by si RNA.Compared with the control group,the m RNA expression of P53 was significantly increased.Flow cytometry showed that compared with the control group,the apoptosis rate of the gliablasts U87 and U251 in the experimental group increased,the cell cycle was blocked in the G2 / M phase,the mitochondrial membrane potential of the cells was significantly reduced,and the intracellular ROS increased significantly.Western blot and Immunofluorescence test showed no significant changes in total caspase3 in the FTL-silenced group,increased expression of clevered-caspase3,Bax,P53,decreased expression of cyclin D1,Bcl-2,P62,and the expression ratio of LC3 I / II increase.Compared with the control group,the positive rate of the FTL silencing group was significantly increased in the TUNNEL test.Results: In glioblastoma cell U87 and U251,silencing FTL expression can cause the cell mitochondrial membrane potential to decrease and increase intracellular reactive oxygen species to increase glioblastoma cell apoptosis and autophagy.At the same time,FTL silencing can cause glioblastoma cells to block in the G2 / M phase,and the above changes may be related to the activation of the P53 pathway.