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. 2022 Jan 20;159(1):6.
doi: 10.1186/s41065-022-00225-0.

SLC3A2 inhibits ferroptosis in laryngeal carcinoma via mTOR pathway

Fangxing Wu  1 , Gaoyun Xiong  1 , Zejun Chen  2 , Chenyang Lei  1 , Qianqian Liu  3 , Yundan Bai  4
Affiliations

SLC3A2 inhibits ferroptosis in laryngeal carcinoma via mTOR pathway

Fangxing Wu et al. Hereditas. .

Abstract

Objective: This study aimed to explore the mRNA and protein expression of SLC3A2 in laryngeal carcinoma cells and tissues, and functional regulatory mechanism of SLC3A2 in cell ferroptosis of laryngeal carcinoma VSports手机版. .

Methods: We chose the key gene-SLC3A2 of DEGs from TCGA by bioinformatics analysis, and then we constructed stable knockdown of SLC3A2 in laryngeal carcinoma cells. MTT assay and clonogenic assay were used to determine cell viability and cell growth, respectively. The mRNA and protein expression were determined by RT-qPCR and western blotting, respectively V体育安卓版. Xenograft tumor model was used to determine the role of SLC3A2 in tumor growth. .

Results: The results of limma analysis recovered that 92 genes were involved in both upregulated DEGs and high risk of poor prognosis, whereas 36 genes were involved in both downregulated DEGs and low risk of poor prognosis. Pathway enrichment analysis indicated that mTOR signaling pathway and ferroptosis exerted a role in regulating these intersection genes V体育ios版. Moreover, SLC3A2 is a key gene in ferroptosis in laryngeal carcinoma. SLC3A2 is highly expressed in laryngeal carcinoma tissues and cells. Patients with high SLC3A2 expression exerted poor survival. SLC3A2 deficiency inhibited cell proliferation and foci formation. Furthermore, knockdown of SLC3A2 expression induced the efficacy of ferroptosis and suppressed ferroptosis related proteins expression. Mechanically, SLC3A2 deficiency facilitated ferroptosis through upregulating the expression of mTOR and P70S6K, whereas inhibited p-mTOR and p-P70S6K expression in laryngeal carcinoma cells. SLC3A2 deficiency inhibited tumorigenesis in nude mice. .

Conclusion: Our study suggests that SLC3A2 negatively regulates ferroptosis through mTOR pathway in laryngeal carcinoma. VSports最新版本.

Keywords: Cell proliferation; Ferroptosis; Laryngeal carcinoma; SLC3A2; mTOR V体育平台登录. .

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Conflict of interest statement

No conflicts of interest.

"V体育平台登录" Figures

Fig. 1
Fig. 1
Screening of DEGs associated with prognosis in laryngeal carcinoma. A TCGA transcriptome data analysis of significantly expressed differential genes in laryngeal cancer. Blue: Decreased expression genes. Red: Increased expression genes. Gray: Not significant change genes. B Single factor cox regression analysis of DEGs involved in prognosis of laryngeal cancer. C DEGs and prognosis related genes were analyzed by venn plot
Fig. 2
Fig. 2
SLC3A2 is a key gene related to ferroptosis in laryngeal carcinoma. A Progression related DEGs were analyzed by KEGG-GO and enrichment. B Expression (p < 0.001) and prognosis (p < 0.045) of SLC3A2 between normal and tumor specimens in the database from TCGA. C RT-qPCR analysis the expression of SLC3A2 mRNA in normal (n = 30) and tumor (n = 30) specimens. D The expression of SLC3A2 protein in normal and tumor specimens was measured by western blotting. N: normal tissues. T: tumor tissues. n = 30. E IHC analysis of the expression of SLC3A2 in normal (n = 30) and tumor (n = 30) specimens. F&G The mRNA and protein expression of SLC3A2 in normal laryngeal epithelial cells and laryngeal carcinoma cells was measured by RT-qPCR and western blotting, respectively. Error bars represent data from three independent experiments (mean ± SD). *p < 0.05,**p < 0.01
Fig. 3
Fig. 3
SLC3A2 deficiency decreases cell proliferation. A HN13 and HN8 cells were transfected with indicated plasmids, and the mRNA expression of SLC3A2 was determined by RT-qPCR. B The expression of SLC3A2 protein was determined by western blotting. C The cell proliferation in SLC3A2 knockdown HN13 and HN8 cells was tested by the MTT assay. D Clonogenic assay was performed to measure the capacity of foci formation in knockdown of SLC3A2 in HN13 and HN8 cells. Error bars represent data from three independent experiments (mean ± SD). *p < 0.05,**p < 0.01
Fig. 4
Fig. 4
SLC3A2 deficiency induces ferroptosis in HN13. A HN13-shNC and shSLC3A2 cells were treated with DMSO, Fer-1 (2.0 μM), Erastin (10.0 μM), Fer-1 (2.0 μM) + Erastin (2.0 μM), RSL3 (2.0 μM) and Fer-1 (2.0 μM) + RSL3 (2.0 μM), respectively, for 48 h and then cell viability was evaluated by MTT assay. In order to better show the difference of cell viability in different treated groups, we converted the cell viability of control group (treated with DMSO) to 100%, and then compare the cell viability of other groups. B&C The levels of total iron level and Fe2+ in laryngeal cancer cells were determined using iron assay kit. D The level of lipid ROS in laryngeal cancer cells was analyzed using cellular ROS assay kit. E The production of mitochondrial superoxide in laryngeal cancer cells was examined using MitoSOX™ Red Mitochondrial Superoxide Indicator. Results presented represent the means of triplicate experiments ± SEM. **p < 0.01
Fig. 5
Fig. 5
SLC3A2 inhibits ferroptosis partly through mTOR pathway. A SLC3A2-Knockdown laryngeal cancer cells were treated with or without L-Leu (activator of mTOR pathway), the protein expression of mTOR, p-mTOR, P70S6K and p-P70S6K was measured by western blotting. B-F The same cells were treated as in (A), then western blotting was perform to measure the ferroptosis related protein expression. G HN13 cells stably expressing control, SLC3A2 knockdown plasmids were injected subcutaneously into nude mice. Representative images showing xenograft mice tumors at day 28 post subcutaneous injection (n = 3). Tumor sizes were measured and depicted as tumor volume and tumor weight. Results presented represent the means of triplicate experiments ± SEM. **p < 0.01. (H) IHC analysis of SLC3A2 and GPX4 expression in xenograft mice tumors. Scale bars, 100 μm
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"V体育安卓版" References

    1. Galluzzi L, Vitale I, Abrams JM, Alnemri ES, Baehrecke EH, Blagosklonny MV, et al. Molecular definitions of cell death subroutines: recommendations of the nomenclature committee on cell death 2012. Cell Death Differ. 2012;19(1):107–120. doi: 10.1038/cdd.2011.96. - DOI - PMC - PubMed
    1. Thompson CB. Apoptosis in the pathogenesis and treatment of disease. Science. 1995;267(5203):1456–1462. doi: 10.1126/science.7878464. - DOI - PubMed
    1. Dixon SJ, Lemberg KM, Lamprecht MR, Skouta R, Zaitsev EM, Gleason CE, et al. Ferroptosis: an iron-dependent form of nonapoptotic cell death. Cell. 2012;149(5):1060–1072. doi: 10.1016/j.cell.2012.03.042. - DOI - PMC - PubMed
    1. Gao M, Monian P, Quadri N, Ramasamy R, Jiang X. Glutaminolysis and transferrin regulate Ferroptosis. Mol Cell. 2015;59(2):298–308. doi: 10.1016/j.molcel.2015.06.011. - DOI - PMC - PubMed
    1. Yang WS, Stockwell BR. Ferroptosis: death by lipid peroxidation. Trends Cell Biol. 2016;26(3):165–176. doi: 10.1016/j.tcb.2015.10.014. - DOI - PMC - PubMed

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