The protective role of mesencephalic astrocyte-derived neurotrophic factor in endoplasmic reticulum stress in RT4-D6P2T schwannoma sells with the S63del MPZ mutation

Bo Sun^1,2, Hongfen Wang¹, Yanran Li¹, Zhengqing He¹, Fang Cui¹, Fei Yang¹, Xusheng Huang^1,*

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¹ Neurological Department of the First Medical Center, Chinese PLA General Hospital, 100853 Beijing, China

² Geriatric Neurological Department of the Second Medical Center & National Clinical Research Center for Geriatric Diseases, Chinese PLA General Hospital, 100853 Beijing, China

^*Correspondence: lewishhuang301@163.com (Xusheng Huang)
Academic Editor: Graham Pawelec

Front. Biosci. (Landmark Ed) 2022, 27(1), 1; https://doi.org/10.31083/j.fbl2701001

Submitted: 18 September 2021 | Revised: 16 November 2021 | Accepted: 26 November 2021 | Published: 4 January 2022

This is an open access article under the CC BY 4.0 license.

Abstract

Background: Endoplasmic reticulum stress (ERS) occurred in S63del mutant CMT1B mice model, and few drugs has been studied. Mesencephalic astrocyte-derived neurotrophic factor (MANF) can inhibit ERS. This study aimed at investigating the effect of MANF on ERS of RT4-D6P2T schwannoma cells with S63del MPZ Mutation. Methods: Experimental grouping: blank control group, blank control + MANF group, lentivirus group, lentivirus + MANF group, S63del MPZ group, S63del MPZ + MANF group. CCK8 and Annexin-FITC/PI were used to detect cell proliferation and apoptosis. JC-1 was used to detect $\Delta{}$ $\Psi{}$ m. MANF, GRP78 and CHOP mRNA and protein were detected by using RT-qPCR, western blotting and immunofluorescence. ER-Tracker and mito-tracker were used to observe the morphology of endoplasmic reticulum (ER) and mitochondria. Results: Cell proliferation decreased (p $<$ 0.001) and apoptosis increased (p $<$ 0.001) in S63del MPZ group; cell proliferation increased (p = 0.005) and apoptosis decreased (p $<$ 0.001) in S63del MPZ + MANF group. $\Delta{}$ $\Psi{}$ m decreased (p $<$ 0.001), MANF, GRP78, CHOP, ATF6, P-PERK/PERK, P-IRE1/IRE1, Bax and Caspase3 increased (p $<$ 0.001) and Bcl2 decreased (p $<$ 0.001) in S63del MPZ group. MANF, GRP78, CHOP, ATF6, P-PERK/PERK, P-IRE1/IRE1, Bax and Caspase3 decreased (p $<$ 0.001) and Bcl2 increased (p $<$ 0.001) in S63del MPZ group. Conclusions: ERS occurred in RT4-D6P2T cells with S63del MPZ mutation, and MANF exerted protective effect in RT4-D6P2T cells with S63del MPZ mutation.

Keywords

Charcot-Marie-Tooth disease type 1B

Endoplasmic reticulum stress

Unfolded protein response

Mesencephalic astrocyte-derived neurotrophic factor

RT4-D6P2T schwannoma cell

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Fig. 1.

Flow cytometry: Annexin V-FITC/PI double staining. C: control, CM: control + MANF, W: wild type, WM: wild type + MANF, S: S63del MPZ, and SM: S63del MPZ + MANF. 1, 2, 3, 4, 5, 6, 7, 8, 9: The difference between two groups was statistically significant (p $<$ 0.05); n = 3.

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