摘要
目的探讨罗氟司特通过调节Yes相关蛋白(Yes-associated protein,YAP)/具有PDZ基序的转录共激活因子(transcriptional coactivator with PDZ-binding motif,TAZ)信号通路对乳腺癌细胞增殖、凋亡和顺铂耐药性的影响。方法选择乳腺癌MCF-7细胞系,构建相应的顺铂(cis-diaminodichloroplatinum,DDP)耐药细胞。将成功建立的MCF-7耐药细胞株(MCF-7/DDP)分为:MCF-7/DDP组、罗氟司特(10μg/mL)+MCF-7/DDP组、罗氟司特(10μg/mL)+YAP激活剂(PY-60,10μmol/L)+MCF-7/DDP组、罗氟司特(10μg/mL)+YAP抑制剂(verteporfin,VTPF,5μmol/L)+MCF-7/DDP组;CCK-8法检测细胞增殖;TUNEL法检测细胞凋亡;qRT-PCR检测细胞中YAP、TAZ mRNA表达;Western blot检测细胞中YAP、TAZ蛋白表达。结果MCF-7/DDP细胞的耐药指数为5.28,采用10μg/mL罗氟司特处理MCF-7/DDP细胞后,耐药指数逆转为1.76。与MCF-7/DDP组相比,罗氟司特+MCF-7/DDP组细胞增殖能力降低,细胞凋亡率提高(P<0.05)。与罗氟司特+MCF-7/DDP组相比,罗氟司特+YAP激活剂+MCF-7/DDP组细胞增殖能力提高,细胞凋亡率降低(P<0.05);与罗氟司特+YAP激活剂+MCF-7/DDP组相比,罗氟司特+YAP抑制剂+MCF-7/DDP组细胞增殖能力降低,细胞凋亡率提高(P<0.05)。qRT-PCR和Western blot结果显示:罗氟司特可显著降低MCF-7/DDP细胞YAP、TAZ mRNA和蛋白表达水平(P<0.05),且其降低作用可被YAP激活剂逆转(P<0.05)。结论罗氟司特可能通过抑制YAP/TAZ信号通路,抑制MCF-7/DDP细胞增殖,促进细胞凋亡,逆转MCF-7/DDP细胞耐药性。
Objective To investigate the effects of roflumilast on the proliferation,apoptosis and cisplatin resistance of breast cancer cells by regulating Yes-associated protein(YAP)/transcriptional coactivator with PDZ-binding motif(TAZ)signaling pathway.Methods The MCF-7 cell line of breast cancer was selected to construct DDP resistant cells.The successfully established drug-resistant MCF-7 cell lines(MCF-7/DDP)were grouped into:MCF-7/DDP group,roflumilast(10μg/mL)+MCF-7/DDP group,roflumilast(10μg/mL)+YAP activator(PY-60,10μmol/L)+MCF-7/DDP group,and roflumilast(10μg/mL)+YAP inhibitor(verteporfin,VTPF,5μmol/L)+MCF-7/DDP group.CCK-8 method was used to detect cell proliferation.TUNEL method was used to detect cell apoptosis.qRT-PCR was used to detect the expression of YAP and TAZ mRNA in cells.Western blot was used to detect the expression of YAP and TAZ proteins in cells.Results The drug resistance index of MCF-7/DDP cells was 5.28,and after treatment with 10μg/mL roflumilast,the drug resistance index was reversed to 1.76.Compared with the MCF-7/DDP group,the proliferation ability of cells in the roflumilast+MCF-7/DDP group was decreased,and the apoptosis rate was increased(P<0.05).Compared with the roflumilast+MCF-7/DDP group,the cell proliferation ability of the roflumilast+YAP activator+MCF-7/DDP group was increased,and the cell apoptosis rate was decreased(P<0.05).Compared with the roflumilast+YAP activator+MCF-7/DDP group,the cell proliferation ability of the roflumilast+YAP inhibitor+MCF-7/DDP group was decreased,and the cell apoptosis rate was increased(P<0.05).qRT-PCR and Western blot results showed that roflumilast significantly reduced the mRNA and protein expression levels of YAP and TAZ in MCF-7/DDP cells(P<0.05),and its reducing effect could be reversed by YAP activators(P<0.05).Conclusion Roflumilast may inhibit the YAP/TAZ signaling pathway,suppress the proliferation of MCF-7/DDP cells,promote cell apoptosis,and reverse drug resistance in MCF-7/DDPcells.
作者
丁晓玲
包蕊
王馨
DING Xiaoling;BAO Rui;WANGXin(Department of Pharmacy,Qinghai Maternal and Child Health Hospital,Xi'ning,Qinghai 810000,China)
出处
《中国优生与遗传杂志》
2025年第5期1092-1097,共6页
Chinese Journal of Birth Health & Heredity
