摘要
目的探讨Toll样受体3(Toll-like receptor 3,TLR3)对乳腺癌细胞MDA-MB-231耐紫杉醇的影响及其作用机制。方法采用浓度递增诱导法建立耐紫杉醇MDA-MB-231乳腺癌细胞株,通过3-(4,5-二甲基噻唑-2-基)-5-(3-羧甲氧基苯基)-2-(4-磺酸苯基)-2H-四唑[3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium,MTS]试剂盒、细胞流式术和台盼蓝染色检测过表达TLR3对耐药细胞增殖和凋亡率的影响;用荧光显微镜观察过表达TLR3对耐药细胞摄取紫杉醇能力的影响;用实时荧光定量聚合酶链式反应(real-time quantitative polymerase chain reaction,RT-qPCR)和蛋白免疫印记(Western Blotting)检测过表达TLR3对耐药细胞中下游基因表达的影响。结果成功构建的耐紫杉醇MDA-MB-231乳腺癌细胞株中TLR3表达水平显著降低;过表达TLR3可显著恢复紫杉醇对乳腺癌细胞增殖的抑制作用,并增强由紫杉醇诱导的细胞凋亡。机制研究发现,耐药细胞中抗凋亡基因p21的表达显著升高,而过表达TLR3可显著抑制p21的表达。结论乳腺癌细胞对紫杉醇的耐药性与TLR3表达下调有关,上调TLR3可通过抑制抗凋亡基因p21的表达,恢复紫杉醇对乳腺癌细胞增殖和存活的抑制作用,为逆转乳腺癌紫杉醇耐药提供潜在靶点和理论依据。
Objective To investigate the effects of Toll-like receptor 3(TLR3)on the resistance of MDA-MB-231 breast cancer cells to paclitaxel and its underlying mechanisms.Methods Paclitaxel-resistant cells were developed by treating MDA-MB-231 cells with gradually increasing dosages of paclitaxel.3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2Htetrazolium(MTS)kit,flow cytometry and Trypan blue staining were performed to determine the proliferation and apoptosis rate in paclitaxel-resistant cells with TLR3 over-expression.The uptake of paclitaxel by cells was determined using immunofluorescent microscopy.The expression of downstream targets of TLR3 was evaluated by real-time quantitative polymerase chain reaction(RTqPCR)and Western blotting.Results A paclitaxel-resistant cell line was successfully established,and TLR3 expression was found to be decreased in these resistant cells.Over-expression of TLR3 in resistant cells restored the inhibitory effects of paclitaxel on cell proliferation and survival.Mechanistically,the expression of the anti-apoptotic gene p21 was significantly elevated,while overexpression of TLR3 could markedly suppress p21 expression.Conclusion The resistance of breast cancer cells to paclitaxel is associated with the downregulation of TLR3 expression.Enhancing TLR3 expression can restore the inhibitory effect of paclitaxel on breast cancer cells proliferation and survival by down-regulating p21 expression.These findings provides a potential therapeutic target and basis for reversing resistance to paclitaxel in breast cancer.
作者
齐雪静
张引兰
曹辉
赵宇
李坤
QI Xuejing;ZHANG Yinlan;CAO Hui;ZHAO Yu;LI Kun(Department of Pharmacy,the Second Hospital of Nanjing,Nanjing University of Chinese Medicine,Nanjing 210003,China;School of Basic Medicine,Qiqihar Medical University,Qiqihar 161006,China;Department of Pharmacy,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450047,China)
出处
《西北药学杂志》
2025年第4期44-50,共7页
Northwest Pharmaceutical Journal
基金
国家自然科学基金项目(编号:81803357)
黑龙江省省属高等学校基本科研业务费科研项目(编号:2019-KYYWF-1226)。
