Sensitivity study of HT22 in ferroptosis

doi:10.3969/j.issn.1006-7795.2020.01.017

Abstract

Abstract: Objectives To investigate whether HT22 cell line is a good cell model to study the ferroptosis and its molecular mechanisms. Methods HT22 cells were treated with Erastin or RSL3, the putative ferroptosis inducers, followed by the observation of morphology change under microscope. Moreover, cell viability was measured by cell counting kit 8 (CCK-8), and cellular lipid peroxidation was measured by MDA assay and the cellular concentration of Fe²⁺ was measured by Fe assay. In addition, the mRNA expression levels of SLC7A11 and PTGS2 were measured by quantitative real time polymerase chain reaction(qRT-PCR). Results Erastin or RSL3 treatment significantly increased lipid peroxidation in HT22, which lead to ferroptosis. However, Erastin or RSL3 did not alter the Fe²⁺ concentration in HT22 cells. In addition, we found that Erastin or RSL3 transcriptionally upregulated the expressions of SLC7A11 and PTGS2. Conclusion HT22 is a suitable cell model to study ferroptosis and the underlying mechanism of the accumulation of lipid peroxidation during ferroptosis.

Key words: ferroptosis, neurons, lipid peroxidation, Fe²⁺

CLC Number:

Q189

Yang Tianli, Yang Yongfei, Yuan Zengqiang. Sensitivity study of HT22 in ferroptosis[J]. Journal of Capital Medical University, 2020, 41(1): 87-91.

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URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2020.01.017

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2020/V41/I1/87

References

[1] Dixon S J,Lemberg K M,Lamprecht M R,et al. Ferroptosis:an iron-dependent form of nonapoptotic cell death[J]. Cell,2012,149(5):1060-1072.
[2] Yagoda N,von Rechenberg M,Zaganjor E,et al. RAS-RAF-MEK-dependent oxidative cell death involving voltage-dependent anion channels[J]. Nature,2007,447(7146):864-868.
[3] Ye F, Chai W, Xie M, et al. HMGB1 regulates erastin-induced ferroptosis via RAS-JNK/p38 signaling in HL-60/NRAS (Q61L) cells[J]. Am J Cancer Res, 2019, 9(4):730-739.
[4] Li L B, Chai R, Zhang S, et al. Iron exposure and the cellular mechanisms linked to neuron degeneration in adult mice[J]. Cells, 2019, 8(2):198-215.
[5] Matsushita M, Freigang S, Schneider C, et al. T cell lipid peroxidation induces ferroptosis and prevents immunity to infection[J]. J Exp Med, 2015, 212(4):555-568.
[6] Friedmann Angeli J P,Schneider M,Proneth B,et al. Inactivation of the ferroptosis regulator Gpx4 triggers acute renal failure in mice[J]. Nat Cell Biol,2014,16(12):1180-1191.
[7] Adedoyin O, Boddu R, Traylor A, et al. Heme oxygenase-1 mitigates ferroptosis in renal proximal tubule cells[J]. Am J Physiol Renal Physiol, 2018, 314(5):702-714.
[8] 王镜岩,朱圣庚,徐长法,等.生物化学(上册)[M]. 3版.北京:高等教育出版社,2002:96-103.
[9] Sui X, Zhang R, Liu S, et al. RSL3 drives ferroptosis through GPX4 inactivation and ROS production in colorectal cancer[J]. Front Pharmacol, 2018, 9(1):1371-1388.
[10] Wang C, Cai X, Hu W, et al. Investigation of the neuroprotective effects of crocin via antioxidant activities in HT22 cells and in mice with Alzheimer's disease[J]. Int J Mol Med, 2019, 43(2):956-966.
[11] Li Z, Chen X, Lu W, et al. Anti-oxidative stress activity is essential for amanita caesarea mediated neuroprotection on glutamate-induced apoptotic HT22 cells and an alzheimer's disease mouse model[J]. Int J Mol Sci, 2017, 18(8):1623-1636.
[12] Lee D S, Cheong S H. Taurine have neuroprotective activity against oxidative damage-induced HT22 death through heme oxygenase-1 pathway[J]. Adv Exp Med Biol, 2017, 975(Pt 1):159-171.
[13] Yang W S, Stockwell B R. Ferroptosis:death by lipid peroxidation[J]. Trends Cell Biol,2016,26(3):165-176.
[14] Zhang Z, Wu Y, Yuan S, et al. Glutathione peroxidase 4 participates in secondary brain injury through mediating ferroptosis in a rat model of intracerebral hemorrhage[J]. Brain Res, 2018, 1701(1):112-125.
[15] Li Q, Han X, Lan X, et al. Inhibition of neuronal ferroptosis protects hemorrhagic brain[J]. JCI Insight, 2017, 2(7):90777-90796.
[16] Fine J M, Forsberg A C, Stroebel B M, et al. Intranasal deferoxamine affects memory loss, oxidation, and the insulin pathway in the streptozotocin rat model of Alzheimer's disease[J]. J Neurol Sci, 2017, 380(1):164-171.