Glaucocalyxin A regulates proliferation,apoptosis and epithelial mesenchymal transformation of colorectal cancer cells by activating the ATF4/CHOP/CHAC1 signaling pathway
LI Mengsen
SUN Longjun
JING Wei
Abstract：Objective To investigate the regulatory effects of glaucocalyxin A(GLA)on the proliferation,apoptosis and epithelial mesenchymal transformation(EMT)of colorectal cancer(CRC)cells by regulating the activating transcription factor 4(ATF4)/CEBP-homologous protein(CHOP)/glutathione-specific gamma-glutamylcyclotransferase 1(CHAC1)signal pathway.Methods Protein expressions of ATF4/CHOP/CHAC1 in CRC tissues and adjacent benign tissues,human CRC cell lines(SW620,SW480,HCT116),and normal colonic epithelial cells(FHC)were detected by Western blot.HCT116 cells were treated with blank control,5 μmol/L GLA(L-GLA group),10 μmol/L GLA(M-GLA group),20 μmol/L GLA(H-GLA group),1 μmol/L GSK2606414(ATF4/CHOP/CHAC1 signaling pathway inhibitor,GSK group)and 10 μmol/L GLA+1 μmol/L GSK2606414(M-GLA+GSK group).Cell proliferation was detected by CCK-8 assay.Flow cytometry was performed to detect cell cycle progression and apoptosis.Transwell assay was performed to detect cell migration and invasion.Protein expressions of key genes in the ATF4/CHOP/CHAC1 signaling pathways,autophagy proteins,EMT-related proteins and apoptosis proteins were detected by Western blot.Results Compared with those of adjacent benign tissues,protein levels of p-ATF4/ATF4,p-CHOP/CHOP,and CHAC1 in CRC tissues were significantly downregulated.They were also significantly downregulated in SW620,SW480 and HCT116 cells,which were the most pronounced in HCT116 cells.Therefore,HCT116 cells were used in the following experiments.Compared with those treated with blank control,protein levels of p-ATF4/ATF4,p-CHOP/CHOP,CHAC1,Bax,cleaved-Caspase-3,E-cadherin,Beclin1 and LC3-Ⅱ/Ⅰ,cell proportion in G1 phase and apoptotic rate were significantly higher,while the OD value,number of migratory and invasive cells,cell proportion in G2/M phase and protein expressions of Bcl-2,vimentin and Snail were significantly lower in HCT116 cells of L-GLA,M-GLA and H-GLA group(all P＜0.05).The above-mentioned trends were opposite in the GSK group.The treatment of GSK2606414 weakened the inhibitory effect of M-GLA on the malignant behaviors of HCT116 cells.Conclusion GLA may inhibit the proliferation,apoptosis and EMT of CRC cells by activating the ATF4/CHOP/CHAC1 signaling pathway.
Keywords：glaucocalyxin Aactivating transcription factor 4(ATF4)/CEBP-homologous protein(CHOP)/glutathione-specific gamma-glutamylcyclotransferase 1(CHAC1)signal pathway(ATF4/CHOP/CHAC1 signal pathway)colorectal cancerepithelial mesenchymal transformationapopto
Publication Date：2025-02-28
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：7( 195-200,206 )