ERK抑制剂增强多柔比星诱导乳腺癌细胞凋亡的作用  

ERK inhibitors enhance apoptosis induced by doxorubicin in breast cancer cells

作　　者：钱江华[1] 刘亚明 杜家如 刘浩[1] QIAN Jiang-hua;LIU Ya-ming;DU Jia-ru;LIU Hao(Dept of Pharmacy, Bengbu Medical College, Bengbu Anhui 233000, China)

机构地区：[1]蚌埠医学院药学系,安徽蚌埠233000

出　　处：《中国药理学通报》2021年第10期1372-1376,共5页Chinese Pharmacological Bulletin

基　　金：安徽省高校自然科学研究项目(No KJ2018A0226)。

摘　　要：目的探讨抑制MEK/ERK信号通路对多柔比星引起人乳腺癌细胞凋亡的影响,为乳腺癌化疗和降低乳腺癌对多柔比星耐药性提供新的靶点。方法MTT法检测衣霉素和多柔比星对MDA-MB-231细胞的增殖抑制作用;免疫组化法检测乳腺癌组织及正常组织中葡萄糖调节蛋白78的表达;TM(4μmol·L^(-1))和ADM(0.5μmol·L^(-1))处理MDA-MB-231细胞,蛋白免疫印迹法检测GRP78、ERK1/2、pERK1/2的表达;MEK抑制剂PD98059(40μmol·L-1)处理MDAMB-231细胞1 h后再给予ADM,检测PD98059组、ADM联合PD98059组的GRP78、ERK1/2、pE RK1/2的表达;RT-PCR检测GRP78转录水平。结果4μmol·L^(-1) TM和0.5μmol·L^(-1) ADM作用于MDA-MB-231细胞48 h细胞存活率分别是(90.72±3.65)%和(60.44±4.37)%;GRP78在乳腺癌组织中强阳性表达;TM和ADM均上调MDA-MB-231细胞GRP78的表达;PD98059明显增加ADM诱导的细胞凋亡率(45%),同时下调GRP78的表达和阻断ADM对GRP78的上调作用,抑制GRP78的转录。结论抑制MEK/ERK信号通路能增强人乳腺癌细胞MDA-MB-231对ADM引起细胞凋亡的敏感性,抑制非折叠蛋白反应的诱导。Aim To investigate the inhibition of MEK/ERK signaling pathway affecting the sensitivity of human breast cancer cells MDA-MB-231 to adriamycin-induced apoptosis so as to provide a new target and reduce adriamycin resistance for breast cancer chemotherapy.Methods The viability of MDA-MB-231 cells exposed to TM and ADM were detected by MTT assay.The expression of GRP78 was examined by immunohistochemistry on tissue sections.TM(4μmol·L^(-1))and ADM(0.5μmol·L^(-1))were used to treat MDA-MB-231 cells,and Western blot was employed to measure protein GRP78、ERK1/2 and pERK expression.MEK signaling pathway inhibitor PD98059(40μmol·L^(-1))was used to pretreat cells for 1 h before treatment with ADM(0.5μmol·L^(-1)),proteins GRP78,ERK1/2 and pERK expression of PD98059 group,PD98059 combined with ADM group were measued.The transcriptional level of GRP78RT-PCR was detected.Results The survival rate of MDA-MB-231 cells was 90.72%and 60.44%,respectively after exposed to(4μmol·L^(-1))TM and(0.5μmol·L^(-1))ADM for 48 h.GRP78 was expressed at significantly higher levels in breast cancer tissues and cells.TM and ADM both up-regulated the expression of GRP78 in MDA-MB-231 cells.PD98059 markedly increased the ADM-induced apoptosis rate to 45%,while down-regulated GRP78 expression,blocked ADM-induced up-regulation of GRP78,and inhibited GRP78 transcription.Conclusions The inhibition of MEK/ERK signaling pathway can enhance the sensitivity of MDA-MB-231 to ADM-induced apoptosis in human breast cancer cells and inhibit ADM-induced unfolded protein response(UPR).

关 键 词：乳腺癌 ER应激 GRP78 多柔比星 凋亡 MEK通路 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]