肺瘤平膏含药血清对肿瘤-免疫-炎性微环境中PI3K/AKT/NF-κB的多能调节作用

Pluripotent regulation of Fei Liu Ping Ointment-containing serum on PI3K/AKT/NF-κB in tumor-immune-inflammatory microenvironment

目的:研究肺瘤平膏含药血清对肿瘤免疫-炎性微环境中肿瘤细胞与非肿瘤细胞的磷脂酰肌醇3激酶/蛋白激酶B/核转录因子κB(PI3K/AKT/NF-κB)分子调节作用,揭示中医药对肿瘤微环境的多能调节机制。方法:建立LL/2-luc-M38肺癌细胞与树突状细胞或RAW264.7巨噬细胞的共培养体系,肺瘤平膏含药血清干预24 h后,收集LL/2-luc-M38肺癌细胞、树突状细胞、RAW264.7巨噬细胞;Western Blot法与实时定量荧光PCR法分别检测PI3K、AKT、NF-κB蛋白及mRNA的表达情况。结果:免疫共培养环境中:与空白血清比较,肺瘤平膏含药血清对LL/2-luc-M38肺癌细胞NF-κB mRNA及蛋白表达有显著抑制作用(P<0.01);肺瘤平膏含药血清可提高树突状细胞中PI3K mRNA和蛋白表达(P<0.05,P<0.01)。炎性共培养环境中:与空白血清比较,肺瘤平膏含药血清可抑制LL/2-luc-M38细胞中PI3K、NF-κB mRNA与蛋白表达(P<0.01,P<0.05)及AKT mRNA表达(P<0.01);肺瘤平膏含药血清可显著提高RAW264.7巨噬细胞中NF-κB mRNA的表达(P<0.01)。结论:肺瘤平膏含药血清对肿瘤-免疫-炎性微环境中的肿瘤细胞与非肿瘤细胞中PI3K、AKT、NF-κB的表达具有多能调节作用。

Objective: To study the PI3K/AKT/NF-κB molecular regulation mechanism of Fei Liu Ping Ointment(FLP)medicated serum on the tumor immune-inflammatory co-culture models of cancer and non-cancerous components, and reveal the multi-directional molecular mechanism of traditional Chinese medicine regulating tumor microenvironment. Methods: A coculture system of LL/2-luc-M38 lung cancer cells and dendritic cells or RAW264.7 macrophages was established. LL/2-luc-M38lung cancer cells, dendritic cells and RAW264.7 macrophages were collected after FLP-medicated serum intervened for 24 hours.Western Blot and real-time quantitative PCR were used to detect the protein and mRNA expressions of PI3K, AKT and NF-κB. Results: Compared with the blank group, FLP-medicated serum could inhibit the mRNA and protein expressions of NF-κB in LL/2-luc-M38 lung cancer cells in the immune co-culture environment(P<0.01). FLP-medicated serum could increase PI3K,AKT mRNA and PI3K protein expression in dendritic cells(P<0.01, P<0.05). FLP-medicated serum in an inflammatory coculture environment could inhibit the mRNA and protein expressions of PI3K and NF-κB in LL/2-luc-M38 cells(P<0.01, P<0.05).In an inflammatory co-culture environment, FLP-medicated serum could increase the mRNA expression of NF-κB in RAW264.7 macrophages(P<0.01). Conclusion: In the immune-inflammatory co-culture microenvironment models, FLP-medicated serum demonstrates multi-directional regulations on the expression of PI3K/AKT/NF-κB both in the tumor cells and non-tumor cells.