The effect of andrographolide regulating STAT3/GPX4 pathway on proliferation and apoptosis of myeloma cells

ZHAO Lanjun; LI Lianghui; MA Xin; GONG Jiaojiao; ZHENG Chenhui; SHI Lin

doi:10.11958/20252757

Abstract：Objective To investigate the effect of andrographolide(Andro)on the proliferation and apoptosis of myeloma cells by regulating the signal transducer and activator of transcription 3(STAT3)/glutathione peroxidase 4(GPX4)pathway.Methods Human multiple myeloma U266 cells were divided into the myeloma group,the Andro low-concentration group,the Andro medium-concentration group,the Andro high-concentration group,the Stattic(STAT3 inhibitor)group and the Andro high-concentration+Colivelin(STAT3 activator)group.Cell proliferation was detected by CCK-8 assay and clone formation assay.Cell apoptosis was detected by flow cytometry.Mitochondrial morphology was observed by transmission electron microscopy.The levels of mitochondrial membrane potential,ferrous ion(Fe²⁺),4-hydroxynonenal(4-HNE),malondialdehyde(MDA)and reduced glutathione(GSH)in cells were detected.The level of reactive oxygen species(ROS)in cells was detected by 2′,7′-dichlorodihydrofluorescein diacetate(DCFH-DA)fluorescent probe.Western blot assay was used to detect the proteins including proliferating cell nuclear antigen(PCNA),B-cell lymphoma-2(Bcl-2)/Bcl-2 associated X protein(Bax),solute carrier family 7 member 11(SLC7A11),p-STAT3 and GPX4.Results Compared with the myeloma group,the mitochondria of U266 cells showed significant morphological changes in the Andro low-,medium-and high-concentration groups,the OD450 value,clone formation rate,mitochondrial membrane potential,GSH level,and the protein levels of PCNA,Bcl-2/Bax,SLC7A11,p-STAT3/STAT3 and GPX4 were decreased,the cell apoptosis rate,the levels of Fe²⁺,4-HNE,MDA and ROS were increased(P＜0.05).Compared with the myeloma group,the changes in the corresponding indexes of U266 cells showed the same trend as above in the Stattic group(P＜0.05).Colivelin reversed the promoting effect of high-concentration Andro on ferroptosis and apoptosis of U266 cells,as well as the inhibitory effect on cell proliferation.Conclusion Andro may induce ferroptosis by inhibiting the STAT3/GPX4 pathway,thereby inhibiting the proliferation and promoting the apoptosis of multiple myeloma cells.