The protective role of mesencephalic astrocyte-derived neurotrophic factor in endoplasmic reticulum stress in RT4-D6P2T schwannoma sells with the S63del MPZ mutation.
The protective role of mesencephalic astrocyte-derived neurotrophic factor in endoplasmic reticulum stress in RT4-D6P2T schwannoma sells with the S63del MPZ mutation.
复制标题
DOI：
10.31083/j.fbl2701001
复制
发表时间：
2022-01
期刊：
影响因子：
--
通讯作者：
Bo Sun;Hong-fen Wang;Yanran Li;Zhengqing He;F. Cui;Fei Yang;Xusheng Huang
中科院分区：
文献类型：
--
作者：
Bo Sun;Hong-fen Wang;Yanran Li;Zhengqing He;F. Cui;Fei Yang;Xusheng Huang
背景内质网应激（ERS）发生在S63del突变型CMT1B小鼠模型中，目前研究的药物很少。中脑星形细胞源性神经营养因子（MANF）可抑制ERS。本研究旨在探讨MANF对S63del MPZ突变的RT4-D6P2T神经鞘瘤细胞ERS的影响。方法实验组分为：空白对照组、空白对照组+ MANF组、慢病毒组、慢病毒+ MANF组、S63del MPZ组、S63del MPZ + MANF组。CCK8和Annexin-FITC/PI检测细胞增殖和凋亡。JC-1用于检测ΔΨm。采用RT-qPCR、western blotting和免疫荧光法检测MANF、GRP78和CHOP的mRNA和蛋白表达。用ER- tracker和mito-tracker观察内质网（ER）和线粒体形态。结果S63del MPZ组细胞增殖减少（p < 0.001），凋亡增加（p < 0.001）；S63del MPZ + MANF组细胞增殖增加（p = 0.005），凋亡减少（p < 0.001）。S63del MPZ组ΔΨm降低（p < 0.001）， MANF、GRP78、CHOP、ATF6、p -PERK/PERK、p -IRE1/IRE1、Bax、Caspase3升高（p < 0.001）， Bcl2降低（p < 0.001）。S63del MPZ组MANF、GRP78、CHOP、ATF6、p -PERK/PERK、p -IRE1/IRE1、Bax、Caspase3降低（p < 0.001）， Bcl2升高（p < 0.001）。结论S63del MPZ突变的RT4-D6P2T细胞发生ERS， MANF对S63del MPZ突变的RT4-D6P2T细胞具有保护作用。
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 ΔΨm. 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. ΔΨm 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.