健脾化瘀方含药血清调控SREBP1/SLC7A11/GPX4轴促进铁死亡抑制胰腺癌PANC-1细胞增殖、侵袭的机制OA

Objective To investigate the mechanism by which the medicated serum of Jianpi Huayu Formula(JPHYF)inhibits the proliferation and invasion of pancreatic cancer PANC-1 cells,based on its role in regulating the SREBP1/SLC7A11/GPX4 axis to promote ferroptosis.Methods Twenty SD rats were randomly divided into a control group and a drug-administered group(n=10 per group).These rats were intragastrically administered either JPHYF decoction(26.8 g·kg-1·d-1)or an equal volume of normal saline once daily for 7 consecutive days to prepare JPHYF medicated serum and blank control serum,respectively.PANC-1 cells were cultured in vitro.The CCK-8 assay was used to measure changes in cell viability after 24-hour intervention with different concentrations(0%,0.5%,2.5%,5%,10%,15%,20%)of JPHYF medicated serum,and appropriate concentrations were selected for subsequent experiments.Colony formation assay was employed to detect cell proliferation capacity;scratch wound healing assay was applied to assess cell migration ability;Transwell assay was applied to evaluate cell invasion ability;fluorescent probes were used to measure intracellular neutral lipid content,reactive oxygen species(ROS)levels,and lipid peroxidation levels;micro-methods were applied to detect cellular ferrous iron(Fe2+),glutathione(GSH),and malondialdehyde(MDA)levels;RT-qPCR and Western Blot were performed to determine the mRNA and protein expression levels of SREBP1,SLC7A11,GPX4,N-cadherin,E-cadherin,and Vimentin.Results(1)Compared with the blank control group,JPHYF medicated serum at concentrations of 2.5%,5%,10%,15%,and 20%significantly inhibited the survival rate of PANC-1 cells(P＜0.05,P＜0.01)in a concentration-dependent manner.The IC50 value of JPHYF medicated serum was 15.40%.Consequently,concentrations of 5%,10%,and 15%medicated serum were selected as the low-,medium-,and high-dose JPHYF groups for subsequent experiments.(2)Compared with the blank control group,the relative colony formation number,24-hour cell migration rate,and number of invasive cells were significantly reduced in the JPHYF low-,medium-,and high-dose groups and the gemcitabine group(5 μmol·L-1)(P＜0.05,P＜0.01).Compared with the blank control group,the JPHYF low-,medium-,and high-dose groups showed significantly increased ROS,Fe2+,and MDA levels of PANC-1 cells(P＜0.05,P＜0.01),significantly decreased GSH levels(P＜0.01),significantly elevated intracellular lipid peroxidation levels(P＜0.05,P＜0.01),and significantly reduced neutral lipid accumulation(P＜0.05,P＜0.01).Furthermore,the mRNA and protein expression levels of SREBP1,GPX4,SLC7A11,N-cadherin,and Vimentin were significantly decreased(P＜0.05,P＜0.01),while the mRNA and protein expression levels of E-cadherin were significantly increased(P＜0.05,P＜0.01)in the JPHYF treatment groups.Conclusions JPHYF medicated serum can effectively inhibit the proliferation,migration,and invasion of pancreatic cancer PANC-1 cells.Its mechanism of action may be related to the suppression of SREBP1 expression,reduction of intracellular lipid accumulation,and induction of ferroptosis in pancreatic cancer cells.