摘要
[目的]探讨东莨菪内酯调控非小细胞肺癌细胞对吉非替尼耐药的潜在机制。[方法]东莨菪内酯处理或不处理的情况下,CCK-8检测非小细胞肺癌细胞PC和吉非替尼耐药细胞PC/GR并计算吉非替尼半最大抑制浓度(IC_(50))。分子对接分析东莨菪内酯与潜在引起吉非替尼耐药的蛋白的相互作用。通过遗传学方法和免疫印迹分析东莨菪内酯和FGL1对肿瘤细胞中与凋亡、增殖相关蛋白表达的影响。[结果]PC9/GR细胞对吉非替尼的IC_(50)为15.66μmol/L。东莨菪内酯处理后,PC9/GR细胞对吉非替尼的IC_(50)为1.81μmol/L。分子对接发现东莨菪内酯与FGL1的结合自由能最低,为-18.44 kcal/mol,结合位点为T70、F96和K150。敲低FGL1后,PC9/GR细胞对吉非替尼的耐药性显著下降,IC_(50)下降为3.52μmol/L。东莨菪内酯处理后,无论过表达还是敲低FGL1,PC9/GR细胞对吉非替尼的耐药性无显著改变。敲低FGL1后,p-EGFR的表达水平下降,Cleaved-PARP1和Cleaved-caspase3的表达水平上升。[结论]东莨菪内酯通过靶向FGL1激活凋亡通路,逆转了非小细胞肺癌细胞对吉非替尼的耐药性。
[Objective]To explore the potential mechanism of Scopoletin regulating gefitinib resistance in non-small cell lung cancer cells.[Method]CCK-8 was used to detect PC in non-small cell lung cancer cells and Gefitinib resistant cells and to calculate Gefitinib semi-maximum inhibitory concentration(IC_(50))under Scopoletin treatment or no treatment.Molecular doc-king analysis of interactions between scopolamine and proteins potentially causing gefitinib resistance.The effects of scopolam-ine and FGL1 on the expression of proteins related to apoptosis and proliferation in tumor cells were analyzed by genetic meth-ods and Western Blotting.[Result]The IC_(50) of PC9/GR cells against gefitinib was 15.66μmol/L.After Scopoletin treatment,the IC_(50) of PC9/GR cells against gefitinib was 1.81μmol/L,respectively.The binding free energy between scopoletin and FGL1 was-18.44 kcal/mol,and the binding sites were T70,F96 and K150.After FGL1 knockdown,the resistance of PC9/GR cells to gefitinib decreased significantly,and IC_(50) decreased to 3.52μmol/L.After Scopoletin treatment,the resistance of PC9/GR cells to gefitinib did not change significantly,regardless of overexpression or FGL1 knockdown.After FGL1 knockdown,p-EGFR expression levels decreased,Cleaved-PARP1 and Cleaved-caspase3 expression levels increased.[Conclusion]Scopo-letin reversed gefitinib resistance in non-small cell lung cancer cells by activating apoptosis pathway by targeting FGL1.
作者
刘威
张帆
龙永贵
LIU Wei;ZHANG Fan;LONG Yonggui(Department of Thoracic Surgery,Leshan People's Hospital,Leshan 614000,China)
出处
《生物技术》
CAS
2024年第6期755-760,766,共7页
Biotechnology
