银杏双黄酮对未分化型甲状腺癌细胞顺铂耐药性的影响与机制研究  

Effect and mechanism of ginkgetin on cisplatin resistance of anaplastic thyroid cancer cells

作　　者：李显彬[1] 王小东 李春艳 李春香 LI Xianbin;WANG Xiaodong;LI Chunyan;LI Chunxiang(Clinical Laboratory,the First Affiliated Hospital of Qiqihar Medical College,Heilongjiang Qiqihar 161041,China;Department of Nuclear Medicine,the First Affiliated Hospital of Qiqihar Medical College,Heilongjiang Qiqihar 161041,China;Department of Ultrasonography,the First Affiliated Hospital of Qiqihar Medical College,Heilongjiang Qiqihar 161041,China)

机构地区：[1]齐齐哈尔医学院附属第一医院检验科,黑龙江齐齐哈尔161041 [2]齐齐哈尔医学院附属第一医院核医学科,黑龙江齐齐哈尔161041 [3]齐齐哈尔医学院附属第一医院超声科,黑龙江齐齐哈尔161041

出　　处：《现代肿瘤医学》2023年第14期2570-2575,共6页Journal of Modern Oncology

基　　金：黑龙江省卫生健康委科研计划(编号:20210909040360)。

摘　　要：目的:探究银杏双黄酮对未分化型甲状腺癌(ATC)细胞顺铂(DDP)耐药性的影响与分子机制。方法:体外培养人ATC细胞8505C,并构建8505C DDP耐药细胞株(8505C/DDP)。用不同浓度的DDP(1.25、2.5、5、10、20μg/mL)或银杏双黄酮(1.25、2.5、5、10、20μmol/L)处理后,CCK-8法检测DDP、银杏双黄酮对8505C、8505C/DDP细胞的毒性作用以及两者联合对8505C/DDP细胞的毒性作用。将8505C/DDP细胞分为DDP组、DDP+银杏双黄酮组、DDP+AMPK抑制剂组、DDP+银杏双黄酮+AMPK抑制剂组,CCK-8法检测细胞增殖活力;Annexin V-FITC/PI双染法检测细胞凋亡情况;划痕愈合实验检测细胞迁移能力;Transwell实验检测细胞侵袭能力;Western Blot检测AMPK/mTOR信号通路相关蛋白表达。结果:DDP对8505C/DDP细胞的半数抑制浓度(IC_(50))高于8505C细胞(P<0.05);银杏双黄酮对8505C/DDP、8505C细胞的IC_(50)无明显差异(P>0.05);银杏双黄酮可增强DDP对8505C/DDP细胞的敏感性(P<0.05)。与8505C细胞比较,8505C/DDP细胞中p-AMPK/AMPK比值降低,p-mTOR/mTOR比值升高(P<0.05);银杏双黄酮可上调8505C细胞和8505C/DDP细胞中p-AMPK/AMPK比值,降低p-mTOR/mTOR比值(P<0.05)。与DDP组比较,DDP+银杏双黄酮组8505C/DDP细胞增殖活力、划痕愈合率、侵袭数量、p-mTOR/mTOR比值降低,凋亡率、p-AMPK/AMPK比值升高(P<0.05);与DDP+银杏双黄酮组比较,DDP+银杏双黄酮+AMPK抑制剂组细胞增殖活力、划痕愈合率、侵袭数量、p-mTOR/mTOR比值升高,凋亡率、p-AMPK/AMPK比值降低(P<0.05)。结论:银杏双黄酮可降低8505C/DDP细胞对DDP的耐药性,增强DDP对8505C/DDP细胞增殖、迁移和侵袭的抑制作用以及对细胞凋亡的诱导作用,其作用机制可能与激活AMPK/mTOR信号通路有关。Objective:To investigate the effect and molecular mechanism of ginkgetin on cisplatin(DDP)resistance in anaplastic thyroid cancer(ATC)cells.Methods:Human ATC cells 8505C were cultured in vitro and 8505C DDP resistant cell lines(8505C/DDP)were constructed and treated with DDP(1.25,2.5,5,10,20μg/mL)or ginkgetin(1.25,2.5,5,10,20μmol/L).CCK-8 assay was used to detect the toxicity of DDP and ginkgetin on 8505C and 8505C/DDP cells and the toxicity of the combination of DDP and ginkgetin on 8505C/DDP cells.The 8505C/DDP cells were separated into DDP group,DDP+ginkgetin group,DDP+AMPK inhibitor group,and DDP+ginkgetin+AMPK inhibitor group.CCK-8 assay was applied to detect cell proliferative viability.Annexin V-FITC/PI double staining was applied to detect cell apoptosis.Scratch-healing assay was applied to examine cell migration ability.Transwell assay was used to detect cell invasiveness.Western Blot was applied to detect expression of proteins related to AMPK/mTOR signaling pathway.Results:The IC_(50) of DDP on 8505C/DDP cells was higher than that in 8505C cells(P<0.05).There was no obvious difference in IC_(50) of ginkgetin on 8505C/DDP and 8505C cells(P>0.05).Ginkgetin could enhance the sensitivity of DDP to 8505C/DDP cells(P<0.05).Compared with 8505C cells,the p-AMPK/AMPK ratio in 8505C/DDP cells was decreased,while the p-mTOR/mTOR ratio was increased(P<0.05).Ginkgetin could increase p-AMPK/AMPK ratio and decrease p-mTOR/mTOR ratio in 8505C cells and 8505C/DDP cells(P<0.05).Compared with DDP group,the proliferation activity,scratch healing rate,invasion number and p-mTOR/mTOR ratio of 8505C/DDP cells in DDP+ginkgetin group were decreased,while the apoptosis rate and p-AMPK/AMPK ratio were increased(P<0.05).Compared with DDP+ginkgetin group,cell proliferation activity,scratch healing rate,invasion number and p-mTOR/mTOR ratio were increased in DDP+ginkgetin+AMPK inhibitor group,while apoptosis rate and p-AMPK/AMPK ratio were decreased(P<0.05).Conclusion:Ginkgetin can reduce the resistance of 8505C/DDP cells to DDP,

关 键 词：银杏双黄酮 AMPK/mTOR信号通路 未分化型甲状腺癌 顺铂耐药性 

分 类 号：R736.1[医药卫生—肿瘤]