Mechanism of saikosaponin D enhancing temozolomide sensitivity in glioma cells via inducing endoplasmic reticulum stress
- VernacularTitle:柴胡皂苷D诱导内质网应激增强胶质瘤细胞替莫唑胺敏感性的机制
- Author:
Gui-Mei LIU
1
;
Rui ZHENG
;
Xiao-Bin LIU
;
Yong-Xian LIU
;
Ya-Ping WANG
;
Yu-Fu ZHANG
;
Jing ZHANG
;
Xiao-Yan JIN
;
Yu-Si LIU
Author Information
- Keywords: saikosaponin D; temozolomide; glioma; proliferation; apoptosis; autophagy; endoplasmic retic-ulum stress
- From: Chinese Pharmacological Bulletin 2024;40(6):1105-1114
- CountryChina
- Language:Chinese
- Abstract: Aim To investigate the synergistic sensiti-zation effect of saikosaponin D(SSD)combined with temozolomide(TMZ)on glioblastoma cells(GBM)and its molecular mechanism.Methods The sensitiv-ity of RG-2,U251 and LN-428 GBM cell lines to SSD and TMZ was analyzed by CCK-8 method combined with HE staining,and the optimal compatible concen-tration was screened.The effect of HE staining com-bined with Hoechst fluorescence staining on the prolif-eration of GBM cell line was detected by clonal forma-tion experiment.The autophagosome formation of GBM cells was observed by monodansylcadaverine(MDC)staining.The expression and distribution of endoplas-mic reticulum stress-related factors and apoptosis and autophagy proteins were detected by Western blot and ICC.Results The sensitivity order of GBM cells to TMZ was RG-2＞U251＞LN-428.The results of com-bined administration showed the synergistic inhibitory effect of SSD combined with TMZ on proliferation of GBM cell lines,which was confirmed by cell cloning formation experiment.Compared with the TMZ group,Hoechst fluorescence staining showed a significant in-crease in the number of nuclear bright staining in the combined administration group.MDC fluorescence staining showed that there were more dense green parti-cles in the cytoplasm of SSD/TMZ plus group than that of TMZ group.Western blot results showed that com-pared with TMZ group,the expression of ER stress markers GRP78,CHOP,p-PERK and ATF6 signifi-cantly increased in SSD/TMZ group(P＜0.05).The expressions of apoptosis proteins caspase-12,caspase-9,caspase-3,cleaved caspase-3,Bax and autophagy proteins LC3 and Beclin-1 significantly increased(P＜0.05),which were verified by ICC test.Conclusions SSD can cooperate with TMZ to inhibit the prolifera-tion of GBM cells and induce apoptosis and autophagy,and enhance the sensitivity of GBM cells to TMZ by ac-tivating endoplasmic reticulum stress pathway.
