内质网应激通过JNK-c-Jun通路调节人乳腺癌细胞生长、自噬和凋亡的分子机制  

Molecular mechanism of endoplasmic reticulum stress regulating growth,autophagy and apoptosis of human breast cancer cells by JNK-c-Jun pathway

作　　者：孙永强[1] 孔天东[2] 杨雯雯 胡金月[1] 党少华[1] 郑帅[1] 郭东慧 Sun Yongqiang;Kong Tiandong;Yang Wenwen;Hu Jinyue;Dang Shaohua;Zheng Shuai;Guo Donghui(Department of Oncology Surgery,the Third People’s Hospital of Zhengzhou,Tumor Hospital of Henan University,Zhengzhou 450000,China;Department of Internal Medicine-Oncology,the Third People’s Hospital of Zhengzhou,Tumor Hospital of Henan University,Zhengzhou 450000,China)

机构地区：[1]郑州市第三人民医院河南大学肿瘤医院肿瘤外科,450000 [2]郑州市第三人民医院河南大学肿瘤医院肿瘤内科,450000

出　　处：《中国实用医刊》2020年第2期10-15,共6页Chinese Journal of Practical Medicine

基　　金：郑州市科技攻关计划资助项目(20130532)。

摘　　要：目的探讨内质网应激通过抑制JNK-c-Jun通路诱导人乳腺癌细胞自噬和凋亡的机制。方法将不同浓度衣霉素不同时间作用于人乳腺癌细胞,确定最佳作用浓度。细胞分组为内质网激活组、内质网激活+SP600125(JNK抑制剂)组、对照组(未做任何处理的常规培养细胞)。使用四甲基偶氮唑蓝法、单丹磺酰尸胺法和流式细胞术分别检测细胞存活、自噬、凋亡;用Western blot分析内质网应激相关蛋白GRP78、CHOP,JNK-c-Jun信号通路相关蛋白JNK、c-Jun,自噬相关蛋白Beclin1,及凋亡相关蛋白Bax、Caspase-3、Caspase-12的表达水平。结果衣霉素对人乳腺癌细胞的最佳作用浓度为100 ng/ml,作用时间为72 h。在此作用条件下,内质网激活相关因子GRP78和CHOP的表达水平较处理前增加(P均<0.05)。与内质网激活+SP600125组和对照组比较,内质网激活组JNK和c-Jun表达水平均上升(P均<0.05)。对照组与内质网激活+SP600125组比较,内质网激活组细胞生长活性明显受到抑制,MDC荧光强度值及细胞凋亡率均增加(P均<0.05)。同时,与内质网激活+SP600125组和对照组比较,内质网激活组中凋亡关键因子Bax、Caspase-3、Caspase-12被激活表达水平均升高(P均<0.05)。结论内质网应激可能通过激活JNK-c-Jun信号通路,降低乳腺癌细胞存活率,进而诱导乳腺癌细胞自噬并促进细胞凋亡。Objective To explore the mechanism of endoplasmic reticulum (ER) stress inducing autophagy and apoptosis of human breast cancer cells by inhibiting c-Jun N-terminal kinases(JNK) -c-Jun signaling pathway.Methods The optimal concentration of tunicamycin was determined based on its effect at different concentration and time on human breast cancer cells. Cells were divided into ER activation group, ER activation+ SP600125 (JNK depressor) group and control group(routindely cultured cells without specific procedure). Methyl thiazolyl tetrazolium assay, dansylcadaverine assay and flow cytometry were used to detect viability, autophagy and apoptosis of cells. Western blot was applied to analyze ER stress-related proteins 78 kDa glucose regulated protein (GRP78) and CCAAT-enhancer-binding protein homolgous protein (CHOP), JNK-c-Jun signaling pathway-related proteins JNK and c-Jun, autophagy-related proteins Beclin1, and apoptosis-related proteins Bax, caspase-3 and caspase-12 expression levels.Results The optimum concentration of tunicamycin on human breast cancer cells was 100 ng/ml, and the action time was 72 h. Under these conditions, the expression levels of GRP78 and CHOP were significantly higher than those before treatment (all P <0.05). The expression levels of JNK and c-Jun in ER activation group were significantly higher than those in ER activation+ SP600125 group and control group (all P <0.05). Compared with the control group and ER activation+ SP600125 group, the proliferation activity of cells in ER activation group was significantly inhibited, while the fluorescence intensity of MDC and the apoptotic rate were significantly increased (all P <0.05). Meanwhile, the expression levels of key apoptotic factors (Bax, Caspase-3 and Caspase-12) in ER activation group, were significantly higher than those in ER activation+ SP600125 group and control group (all P <0.05).Conclusions ER stress may inhibit the cell growth, further induce autophagy and apoptosis of breast cancer cells by activating JNK-c-Jun signaling pa

关 键 词：内质网 应激 JNK/c-Jun信号通路 细胞自噬 细胞凋亡 

分 类 号：R737[医药卫生—肿瘤]