江苏大学学报（医学版）2024，Vol.34Issue(1)：1-10,10.DOI:10.13312/j.issn.1671-7783.y220273
hnRNPA2B1通过抑制转铁蛋白受体增强胰腺癌细胞对铁死亡的抵抗
hnRNPA2B1 enhances the resistance of pancreatic cancer cells to ferroptosis by inhibiting transferrin receptor
摘要
Abstract
Objective:To investigate the role of heterogeneous nuclear ribonucleoprotein A2B1(hnRNPA2B1)in ferroptosis of pancreatic cancer cells and its potential mechanisms.Methods:Three pancreatic cancer cell lines PaTu8988,MIA-paca2 and PANC1 were treated with different concentrations of Erastin for 3 days.The cell viability was detected by CCK8 method.The half inhibitory concentrations(IC50)of Erastin in three kinds of pancreatic cancer cell lines were calculated and their resistance capabilities to Erastin were compared;qRT-PCR and Western blotting were used to detect the relative expression levels of mRNA and protein of hnRNPA2B1 in pancreatic cancer cells.The differential expression of the hnRNPA2B1 in pancreatic cancer tissues and normal tissues was analyzed by GEPIA,and the survival prognosis of patients with differential expression of hnRNPA2B1 was analyzed from clinical data sets from GEO database.PaTu8988 cells was transfected with sh-Control and sh-hnRNPA2B1 plasmid,respectively,to knock down hnRNPA2B1;PANC1 cells was transfected with Vector and Flag-hnRNPA2B1 plasmid,respectively,to overexpress hnRNPA2B1;and the transfection efficiency was verfied by qRT-PCR and Western blotting,respectively;the effects of knockdown or overexpression of hnRNPA2B1 on the migration and invasion were detected by Transwell assay,and the effects on the proliferation of pancreatic cancer cells were detected by CCK8 assay.The cells in sh-Control,sh-hnRNPA2B1,Vector and Flag-hnRNPA2B1 group were treated with control(0 μmol/L Erastin)and Erastin(IC50 concentration),respectively,then flow cytometry was used to determine the lipid peroxidation level,and colorimetry was used to determine the malindialdehyde(MDA)and tissue iron concentration;in addition to Erastin,the sh-hnRNPA2B1 and Flag-hnRNPA2B1 groups were added with Ferrostatin-1,Necrostatin-1,ZVAD-FMK,respectively,the cells activity was detected by trypan blue dye exclusion.The relative expression levels of RNA and protein of transferrin receptor(TFRC),ferritin heavy chain,glutathione peroxidase 4 and solute carrier family 7 member 11 were detected by qRT-PCR and Western blotting,respectively.Results:The resistance of three types of pancreatic cancer cells to Erastin from high to low were PaTu8988,MIA-paca2 and PANC1,corresponding Erastin IC50 was 18.020,15.760 and 2.947 μmol/L,respectively(P＜0.05).The relative expression level of hnRNPA2B1 was the highest in PaTu8988 cells,followed by MIA-paca2 cells and the lowest in PANC1 cells(P＜0.05),which was the same with Erastin IC50.The expression level of hnRNPA2B1 in pancreatic cancer tissues was significantly higher than that in normol tissues,and the prognosis of patients with higher expression of hnRNPA2B1 was poor(P＜0.05).Down-regulating the expression of hnRNPA2B1,the migration,invasion and proliferation ability of PaTu8988 cells were greatly reduced,while up-regulating was the opposite(P＜0.05).Erastin-induced cell death was significantly restored by Ferrostatin-1 not by ZVAD-FMK or Necrostatin-1(P＜0.05);compared with sh-Control group,the level of lipid peroxidation,MDA and iron concentration in sh-hnRNPA2B1 group were significantly increased(P＜0.05);compared with Vector group,Flag-hnRNPA2B1 group showed lower level of lipid peroxidation,MDA and iron concentration(P＜0.05).Upregulation of hnRNPA2B1 inhibited the expression of TFRC,while interference with hnRNPA2B1 produced the opposite effect(P＜0.05).Conclusion:hnRNPA2B1 could inhibit the intracelluar accumulation of iron by inhibiting the expression of TFRC,thus promoting the resistance of pancreatic cancer cells to ferroptosis.关键词
hnRNPA2B1/胰腺癌/铁死亡/铁代谢/转铁蛋白受体Key words
hnRNPA2B1/pancreatic cancer/ferroptosis/metabolism of iron/transferrin receptor(TFRC)分类
医药卫生引用本文复制引用
李政,刘雅雯,陈家希,王慧之,于正悦,王舜宇,龚爱华,徐岷..hnRNPA2B1通过抑制转铁蛋白受体增强胰腺癌细胞对铁死亡的抵抗[J].江苏大学学报（医学版）,2024,34(1):1-10,10.基金项目
国家自然科学基金面上项目(82072754) （82072754）
江苏省卫生健康委员会面上项目(M2020011) （M2020011）
江苏省重点研发计划社会发展项目(BE2018689) （BE2018689）
镇江市重点研发计划社会发展项目(SH2018033) （SH2018033）
