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

Abstract
Figure/Table
References (0)
Related Citation (15)

Download: PDF (2230 KB) HTML (1 KB)
Export: BibTeX | EndNote (RIS) Supporting Info

Guide

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 weredetected 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 ironconcentration (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 t he intracelluar accumulation of iron by inhibiting the expression of TFRC, thus promoting the resistance of pancreatic cancer cells to ferroptosis.

Key words：

Received: 30 December 2022

Fund:

	Service

	E-mail this article
	Add to my bookshelf
	Add to citation manager
	E-mail Alert
	RSS
	Articles by authors
	LI Zheng1
	LIU Yawen1
	CHEN Jiaxi2
	WANG Huizhi1
	YU Zhengyue1
	WANG Shunyu1
	GONG Aihua1
	XU Min3

Cite this article:

LI Zheng1,LIU Yawen1,CHEN Jiaxi2等. hnRNPA2B1 enhances the resistance of pancreatic cancer cells to ferroptosis by inhibiting transferrin receptor[J]. Journal of Jiangsu University(Medicine Edition), 2024, 24(01): 1-10.

URL:

http://zzs.ujs.edu.cn/xbyxb/EN/ OR http://zzs.ujs.edu.cn/xbyxb/EN/Y2024/V24/I01/1

［1］Tao JX, Yang G, Zhou WC, et al. Targeting hypoxic tumor microenvironment in pancreatic cancer［J］. J Hematol Oncol, 2021, 14(1): 14.

［2］Luo GP, Jin KZ, Deng SM, et al. Roles of CA199 in pancreatic cancer: Biomarker, predictor and promoter［J］. Biochim Biophys Acta Rev Cancer, 2021, 1875(2): 188409.

［3］Kajioka H, Kagawa S, Ito A, et al. Targeting neutrophil extracellular traps with thrombomodulin prevents pancreatic cancer metastasis［J］. Cancer Lett, 2021, 497: 1-13.

［4］Yu S, Zhang CY, Xie KP. Therapeutic resistance of pancreatic cancer: Roadmap to its reversal［J］. Biochim Biophys Acta Rev Cancer, 2021, 1875(1): 188461.

［5］Chen X, Kang R, Kroemer G, et al. Ferroptosis in infection, inflammation, and immunity［J］. J Exp Med, 2021, 218(6): e20210518.

［6］Chen X, Kang R, Kroemer G, et al. Broadening horizons: the role of ferroptosis in cancer［J］. Nat Rev Clin Oncol, 2021, 18(5): 280-296.

［7］蔡晓杰, 王鸣, 高洁, 等. 二甲双胍逆转低氧诱导的胰腺癌细胞铁死亡抵抗及潜在的作用机制［J］. 江苏大学学报(医学版), 2020, 30(2): 125-130.

［8］Xiao C, Fu X, Wang YT, et al. Transferrin receptor regulates malignancies and the stemness of hepatocellular carcinomaderived cancer stemlike cells by affecting iron accumulation［J］. PLoS One, 2020, 15(12): e0243812.

［9］Bao ZH, Hou XB, Li HL, et al. The mechanism and progress of ferroptosis in pancreatic cancer［J］. Acta Histochem, 2022, 124(6): 151919.

［10］Manz DH, Blanchette NL, Paul BT, et al. Iron and cancer: recent insights［J］. Ann N Y Acad Sci, 2016, 1368(1): 149-161.

［11］Jiang FJ, Tang XZ, Tang C, et al. HNRNPA2B1 promotes multiple myeloma progression by increasing AKT3 expression via m6Adependent stabilization of ILF3 mRNA［J］. J Hematol Oncol, 2021, 14(1): 54.

［12］Wang L, Wen MY, Cao XT. Nuclear hnRNPA2B1 initiates and amplifies the innate immune response to DNA viruses［J］. Science, 2019, 365(6454): eaav0758.

［13］Tang JZ, Chen ZM, Wang Q, et al. hnRNPA2B1 promotes colon cancer progression via the MAPK pathway［J］. Front Genet, 2021, 12: 666451.

［14］Chen ZY, Chen X, Lei TY, et al. Integrative analysis of NSCLC identifies LINC01234 as an oncogenic lncRNA that interacts with HNRNPA2B1 and regulates miR106b biogenesis［J］. Mol Ther, 2020, 28(6): 1479-1493.

［15］Chen CH, Luo YM, He W, et al. Exosomal long noncoding RNA LNMAT2 promotes lymphatic metastasis in bladder cancer［J］. J Clin Invest, 2020, 130(1): 404-421.

［16］Dai SJ, Zhang J, Huang S, et al. HNRNPA2B1 regulates the epithelialmesenchymal transition in pancreatic cancer cells through the ERK/snail signalling pathway［J］. Cancer Cell Int, 2017, 17(1): 12.

［17］Chen ZY, Cai L, Zhu J, et al. Fyn requires HnRNPA2B1 and Sam68 to synergistically regulate apoptosis in pancreatic cancer［J］. Carcinogenesis, 2011, 32(10): 1419-1426.

［18］Tang FJ, Chen L, Gao H, et al. m6A: An emerging role in programmed cell death ［J］. Front Cell Dev Biol, 2022, 10: 817112.

［19］Zhao YC, Li YQ, Zhang RF, et al. The role of erastin in ferroptosis and its prospects in cancer therapy［J］. Onco Targets Ther, 2020, 13: 5429-5441.

［20］Liu Y, Shi SL. The roles of hnRNP A2/B1 in RNA biology and disease［J］. Wiley Interdiscip Rev RNA, 2021, 12(2): e1612.

［21］Pan ZW, Zhao RR, Li BY, et al. EWSR1induced circNEIL3 promotes glioma progression and exosomemediated macrophage immunosuppressive polarization via stabilizing IGF2BP3［J］. Mol cancer, 2022, 21(1): 6.

［22］Forciniti S, Greco L, Grizzi F, et al. Iron metabolism in cancer progression［J］. Int J Mol Sci, 2020, 21(6): 2257.

［23］Zhang C, Liu XY, Jin SD, et al. Ferroptosis in cancer therapy: a novel approach to reversing drug resistance［J］. Mol Cancer, 2022, 21(1): 47.

［24］Brown CW, Amante JJ, Chhoy P, et al. Prominin2 drives ferroptosis resistance by stimulating iron export［J］. Dev Cell, 2019, 51(5): 575-586.

［25］Liu YW, Huang P, Li Z, et al. Vitamin C sensitizes pancreatic cancer cells to Erastininduced ferroptosis by activating the AMPK/Nrf2/HMOX1 pathway ［J］. Oxid Med Cell Longev, 2022, 15: 5361241.

［26］Guo SS, Chen YX, Xue XF, et al. TRIB2 desensitizes ferroptosis via βTrCPmediated TFRC ubiquitiantion in liver cancer cells［J］. Cell Death Discov, 2021, 7(1): 196.

［27］Lu YX, Yang Q, Su YB, et al. MYCN mediates TFRCdependent ferroptosis and reveals vulnerabilities in neuroblastoma［J］. Cell Death Dis, 2021, 12(6): 511.

［28］Zhang J, Chen SJ, Wei ST, et al. CircRAPGEF5 interacts with RBFOX2 to confer ferroptosis resistance by modulating alternative splicing of TFRC in endometrial cancer［J］. Redox Biol, 2022, 57: 102493.