松果菊苷调节受体相互作用蛋白1/受体相互作用蛋白3/混合谱系激酶结构域样蛋白信号通路对肺癌细胞化疗敏感性的影响  

Impact of echinacoside on chemosensitivity of lung cancer cells by regulating the RIP1/RIP3/MLKL signal pathway

作　　者：张晋源 ZHANG Jinyuan(Department of Oncology,Shanxi Provincial Hospital,Taiyuan,Shanxi 030001)

机构地区：[1]山西省人民医院肿瘤科,山西太原030001

出　　处：《河北中医》2025年第4期606-611,共6页Hebei Journal of Traditional Chinese Medicine

摘　　要：目的探讨松果菊苷(ECH)调节受体相互作用蛋白1(RIP1)/RIP3/混合谱系激酶结构域样蛋白(MLKL)信号通路对肺癌细胞化疗敏感性的影响。方法将A549/DDP细胞分为A549组(未处理的A549/DDP细胞)、L-ECH组(5μM ECH处理A549/DDP细胞)、M-ECH组(10μM ECH处理A549/DDP细胞)、H-ECH组(20μmol/L ECH处理A549/DDP细胞)、GSK2982772组(16 nM RIP1/RIP3/MLKL通路抑制剂GSK2982772处理A549/DDP细胞)、H-ECH+GSK2982772组(20μM ECH和16 nM RIP1/RIP3/MLKL通路抑制剂GSK2982772共同处理A549/DDP细胞);用不同浓度顺铂(0.5、1.0、1.5、2.0、2.5、5.0μg/mL)处理A549以及A549/DDP细胞检测化学敏感性;MTT法检测A549/DDP细胞活力;流式细胞术检测A549/DDP细胞凋亡;Transwell检测A549/DDP细胞侵袭、迁移能力;Western blot检测EMT蛋白、自噬蛋白以及RIP1/RIP3/MLKL信号通路蛋白表达。结果A549细胞IC50值(1.38μg/mL)显著低于A549/DDP细胞IC50值(2.42μg/mL),因此,选用2.5μg/mL顺铂做后续实验。与A549组相比,L-ECH组、M-ECH组、H-ECH组OD值、A549/DDP细胞迁移、侵袭数量、vimentin、Snail蛋白水平依次显著下降(P<0.05),凋亡率、Beclin1、LC3-II/I、RIP1、RIP3、MLKL、E-cadherin蛋白水平依次显著升高(P<0.05),呈现剂量依赖性,而GSK2982772组以上指标趋势相反;与H-ECH组相比,H-ECH+GSK2982772组OD值、A549/DDP细胞迁移、侵袭数量、vimentin、Snail蛋白水平显著增加(P<0.05),凋亡率、Beclin1、LC3-II/I、RIP1、RIP3、MLKL、E-cadherin蛋白水平显著减少(P<0.05)。结论ECH可能通过激活RIP1/RIP3/MLKL信号通路对肺癌细胞化疗敏感性产生影响。Objective To investigate the impact of echinacoside(ECH)on chemosensitivity of lung cancer cells by regulating the Receptor-Interacting Protein 1(RIP1)/RIP3/Mixed Lineage Kinase Domain-Like(MLKL)signaling pathway.Methods A549/DDP cells were treated with blank control,low-dose(5μM),medium-dose(10μmol/L)and high-dose ECH(20μmol/L),16 nM GSK2982772(the RIP1/RIP3/MLKL pathway inhibitor)and high-dose ECH+GSK2982772.A549 and A549/DDP cells were treated with cisplatin at the concentrations of 0.5,1.0,1.5,2.0,2.5,and 5.0μg/ml to detect the sensitivity to chemotherapy.Cell viability and apoptosis of A549/DDP cells were detected by microwave thermoacoustic tomography(MTT)assay,and flow cytometry,respectively.Transwell was applied to detect the invasion and migration abilities of A549/DDP cells,and Western blot was applied to detect the expressions of epithelial-mesenchymal transition(EMT)proteins,autophagy proteins and proteins in the RIP1/RIP3/MLKL signaling pathway.Results The IC50 value of A549 cells(1.38μg/mL)was significantly lower than that of A549/DDP cells(2.42μg/mL),and therefore,2.5μg/ml cisplatin was selected for the following experiments.Compared with A549/DDP cells treated with blank control,those treated with low-dose,medium-dose and high-dose ECH showed significantly lower optical density(OD),migratory and invasive cell numbers and protein levels of Vimentin and Snail,but higher apoptotic rate and protein levels of Beclin 1,LC3-II/I,RIP1,RIP3,MLKL,and E-cadherin in a dose-dependent manner(P<0.05).Treatment of GSK2982772 showed the opposite results.Compared with A549/DDP cells treated with high-dose ECH,those treated with high-dose ECH+GSK2982772 showed significantly higher OD,migratory and invasive cell numbers and protein levels of Vimentin and Snail,but lower apoptotic rate and protein levels of Beclin 1,LC3-II/I,RIP1,RIP3,MLKL,and E-cadherin(P<0.05).Conclusion ECH may affect the chemosensitivity of lung cancer cells by activating the RIP1/RIP3/MLKL signaling pathway.

关 键 词：松果菊苷 RIP1/RIP3/MLKL信号通路 肺癌 化疗敏感性 

分 类 号：R730.5[医药卫生—肿瘤]