肺积方调控补体相关蛋白CFHR5/MBL2/C9抑制肺癌细胞迁移、侵袭及动物模型肺转移  

Feiji Formula inhibits the migration and invasion of lung cancer cells and lung metastasis in animal models by regulating complement-related proteins CFHR5/MBL2/C9

作　　者：罗斌[1,3] 王衍鸿 刘佳君 刘诗卉 陆鑫熠 李佳轩 阙祖俊 田建辉[1,2] LUO Bin;WANG Yanhong;LIU Jiajuna;LIU Shihui;LU Xinyi;LI Jiaxuan;QUE Zujun;TIAN Jianhui(a.Clinical Oncology Center,Shanghai University of Traditional Chinese Medicine,Shanghai 200071,China;Cancer Research Institute,Shanghai Municipal Hospital of Traditional Chinese Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200071,China;Department of Oncology,Longhua Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China)

机构地区：[1]上海中医药大学附属市中医医院肿瘤临床医学中心,上海200071 [2]上海中医药大学附属市中医医院肿瘤研究所,上海200071 [3]上海中医药大学附属龙华医院肿瘤科,上海200032

出　　处：《中国肿瘤生物治疗杂志》2024年第12期1194-1203,共10页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金青年项目(No.82104943);国家自然科学基金面上项目(No.82174245);上海市卫生健康委员会领军人才基金(No.2022LJ014);上海市炎癌转化病证生物学前沿研究基地项目(No.2021科技03-12);岐黄工程·第五批全国中医临床优秀人才项目[No.国中医药人教函(2022)239号]。

摘　　要：目的:探讨肺积方对肺癌细胞迁移、侵袭及在动物模型中肺转移的影响及其可能的机制。方法:利用TNMplot数据库、TCGA数据库和DAVID数据库,通过网络药理学方法分析补体相关蛋白CFHR5/MBL2/C9在肺转移瘤组织中的表达及其与免疫细胞浸润的关系,以及相关的生物过程和信号通路。建立2LL细胞小鼠皮下移植瘤模型,用2 g/mL肺积方汤液灌胃给药,每次0.2 mL,连续干预21 d,观察肺积方对模型小鼠肺转移发生率、肺转移灶数目及肺组织CFHR5/MBL2/C9蛋白表达的影响。外泌体示踪实验观察CTC-TJH-01和H1299细胞分泌和吞噬外泌体的能力。采用不同质量浓度的肺积方冻干粉溶液处理H1299和CTC-TJH-01细胞,采用CCK-8、划痕愈合实验、Transwell实验和WB法检测肺积方对肺癌细胞活力、侵袭、迁移和CFHR5/MBL2/C9蛋白表达的影响。结果:网络药理学分析结果提示,CFHR5/MBL2/C9蛋白在肺转移组织中呈高表达(P<0.05)且与调控免疫应答中的补体系统密切相关。与对照组相比,肺积方组模型小鼠的肺转移灶数量显著减少(P<0.05)。0~200μg/mL肺积方对H1299和CTC-TJH-01细胞的活力均无显著影响(均P>0.05)。CTC-TJH-01和H1299细胞均可分泌并吞噬对方的外泌体。与0μg/mL肺积方对照组相比,50~200μg/mL肺积方组H1299和CTC-TJH-01细胞的迁移和侵袭能力均显著下降(P<0.05或P<0.01),细胞中的CFHR5和MBL2蛋白表达水平均显著降低(P<0.05或P<0.01),其中CTC-TJH-01细胞在200μg/mL肺积方溶液处理后C9蛋白表达水平升高(P<0.05)。结论:肺积方能够通过调控补体相关蛋白CFHR5/MBL2/C9抑制肺癌细胞的迁移和侵袭及模型小鼠的肺转移,这可能与外泌体介导的细胞间通信有关。Objective:To investigate the effects of Feiji Formula on the migration and invasion of lung cancer cells,as well as lung metastasis in animal models and explore its possible mechanisms.Methods:TNMplot,TCGA,and DAVID databases were used to analyze the expression of complement-related proteins(CFHR5[complement factor H-related protein 5],MBL2[mannose-binding lectin 2],C9[complement component 9])in lung metastatic tissues and their relationship with immune cell infiltration,as well as related biological processes and signaling pathways.A subcutaneous xenograft mice model was established using 2LL cells.Mice were administered 2 g/mL Feiji Formula decoction(0.2 mL per dose)via oral gavage for 21 days.The effects of Feiji Formula on the incidence of lung metastasis,the number of lung metastatic nodules,and the protein expression of CFHR5/MBL2/C9 in the lung tissues of the model mice were observed.Exosome tracing assay was performed to observe the secretion and uptake of exosomes by CTC-TJH-01 and H1299 cells.Different concentrations of Feiji Formula were applied to treat H1299 and CTC-TJH-01 cells,and its effects on cell viability,invasion,migration,and CFHR5/MBL2/C9 protein expression were detected by CCK-8,scratch healing assay,Transwell assay,and WB method.Results:Network pharmacology analysis showed that CFHR5/MBL2/C9 proteins were highly expressed in lung metastatic tissues(all P<0.05)and were closely related to the complement system involved in immune response regulation.Compared with the control group,the Feiji Formula group demonstrated a significant reduction in the number of lung metastatic nodules(P<0.05).Feiji Formula(0-200μg/mL)had no significant effect on the viability of H1299 and CTC-TJH-01 cells(both P>0.05).Both CTC-TJH-01 and H1299 cells could secrete and uptake each other's exosomes.Compared with the 0μg/mL control group,Feiji Formula at concentrations of 50-200μg/mL significantly inhibited the migration and invasion abilities of H1299 and CTC-TJH-01 cells(P<0.05 or P<0.01),and significantly redu

关 键 词：肺积方 肺癌 转移 外泌体蛋白 补体系统 

分 类 号：R734.2[医药卫生—肿瘤] R730.52[医药卫生—临床医学] R285.5