Background:Mesencephalic astrocyte-derived neurotrophic factor(MANF)is a new candidate growth factor for dopaminergic neurons against endoplasmic reticulum stress(ER stress).HSP70 family,a chaperon like heat shock protein family,was proved to be involved in the MANF induced survival pathway in 6-OHDA treated SHSY-5Y cells.However,the ER stress relative transcriptome,in MANF signaling cascades is still investigated.The involvement of HSP70,a 70kd member of HSP70 family,need further to be verified.Methods:The cell apoptosis was assayed by MTT,TUNEL staining and western blot of cleaved Caspase-3.The differentially expressed genes in SHSY-5Y cells under different conditions(control,6-OHDA,6-OHDA+MANF)were investigated by RNA-seq.Expression of HSP70 was further confirmed by real-time PCR.RNAi knockdown for HSP70 was performed to investigate the role of HSP70 in the MANF signaling pathway.Results:MANF inhibits 6-OHDA-induced apoptosis in SHSY-5Y cells.Six ER stress relative genes(HSP70,GRP78,xbp-1,ATF-4,ATF-6,MAPK)were found enriched in 6-OHDA+MANF treatment group.HSP70 was the most significantly up-regulated gene under 6-OHDA+MANF treatment in SHSY-5Y cells.RNAi knockdown for HSP70 inhibits the protective effects of MANF against 6-OHDA toxicity in SHSY-5Y cells.Conclusion:MANF exerts a protective role against 6-OHDA induced apoptosis in SHSY-5Y cells via up-regulating some ER stress genes,including HSP70 family members.The HSP70 expression level plays a key role in MANFmediated survival pathway.
