粉防己碱通过NF-κB通路抑制肿瘤相关成纤维细胞分泌IL-8阻断肺腺癌细胞的侵袭转移  

Tetrandrine Inhibits Invasion and Metastasis of Lung Adenocarcinoma Cells by Inhibiting IL-8 Secretion in Tumor-Associated Fibroblasts via NF-κB Pathway

作　　者：万海军[1] 邱水玮 陈霖[1] 于进江 吕定量[1] WAN Hai-jun;QIU Shui-wei;CHEN Lin;YU Jin-jiang;LV Ding-liang(People’s Hospital of Quzhou,Quzhou 324000,China)

机构地区：[1]衢州市人民医院,浙江衢州324000

出　　处：《肿瘤学杂志》2020年第5期402-406,共5页Journal of Chinese Oncology

基　　金：浙江省中医药科学研究基金(2015ZA216);衢州市指导性科技项目(2016068);衢州市指导性科技项目(2018071)。

摘　　要：[目的]研究粉防己碱(tetrandrine,Tet)通过调控肿瘤相关成纤维细胞(CAF)旁分泌因子的表达抑制肺腺癌细胞侵袭转移的作用效应,探讨NF-κB通路在Tet抑制CAF分泌IL-8促进肺腺癌细胞侵袭转移中的重要作用。[方法]采用条件培养基(CM)刺激胚肺成纤维细胞MRC5建立CAF模型,Western blot实验分析Tet作用前后NF-κB通路蛋白IKK、IκB和p65表达和磷酸化水平,ELISA实验检测Tet作用前后CAF分泌IL-8情况,以及Tet对CAF诱导肺腺癌细胞H1975侵袭转移的影响。抗体封闭培养基中游离的IL-8,分析CAF诱导H1975细胞侵袭转移能力的变化,验证IL-8是否Tet调控CAF旁分泌因子的表达抑制H1975细胞侵袭转移的关键因子。[结果] Tet作用24h,CAF中NF-κB通路蛋白IKK、IκB和p65蛋白的磷酸化水平显著性下降;Tet组培养基上清液中IL-8含量为(151.7±24.8)pg/ml,显著性低于模型组的(384.7±39.5)pg/ml(P<0.001);模型组诱导H1975细胞迁移的数量为141.7±21.5,Tet组为41.3±11.9(P<0.01);模型组诱导H1975细胞侵袭的数量为129.7±25.1,Tet组为56.7±14.5(P<0.05)。模型组诱导H1975细胞迁移的数量为152.3±25.5,IL-8封闭(aIL-8)组为61.7±15.5(P<0.01);模型组诱导H1975细胞侵袭的数量为136.3±14.5,aIL-8组为73.3±12.7(P<0.01)。[结论] Tet具有抑制CAF诱导肺腺癌细胞侵袭转移的作用效应,其机制与抑制NF-κB通路的活性和CAF分泌IL-8有关。[Objective] To investigate the inhibitory effect of tetrandrine(Tet) on the invasion and metastasis of lung adenocarcinoma cells and its mechanisms. [Methods] The conditioned medium(CM) was used to stimulate embryonic lung fibroblasts MRC5 to establish a cancer-associated fibroblast(CAF) model.Western blot was used to analyze the expression and phosphorylation level of NF-κB pathway proteins IKK,IκB and p65 before and after Tet treatment. ELISA assay was used to assess the secretion of IL-8 in CAF,and the ability of CAF to induce the invasion and metastasis of human lung adenocarcinoma H1975 cells was analyzed with Transwell assay. The free IL-8 in the culture medium was blocked by antibody(aIL-8 group),and the change of CAF induced effect was evaluated in H1975 cells. [Results] After 24 h of Tet treatment,the phosphorylation levels of NF-κB pathway proteins IKK,IκB,and p65 protein in CAF decreased significantly. The content of IL-8 in the supernatant of the Tet group was(151.7±24.8) pg/ml,which was significantly lower than the model group(384.7±39.5) pg/ml(P<0.001). The number of H1975 cell migration in the model group was 141.7 ±21.5,and that in Tet group was 41.3 ±11.9(P <0.01). The number of H1975 cell invasion in the model group was 129.7±25.1,and that in Tet group was 56.7±14.5(P<0.05). The number of H1975 cell migration induced by the model group was 152.3 ±25.5,and that of the IL-8 blocked(aIL-8) group was 61.7±15.5(P<0.01). The number of H1975 cells in the model group was136.3±14.5,and It was 73.3±12.7 in the aIL-8 group(P<0.01). [Conclusion] Tetrandrine can inhibit the CAF-induced invasion and metastasis of lung adenocarcinoma cells,which is associated with the inhibition of the activity of NF-κB pathway and the secretion of IL-8 by CAF.

关 键 词：粉防己碱 NF-ΚB通路 肿瘤相关成纤维细胞 肺腺癌 侵袭转移 

分 类 号：R734.2[医药卫生—肿瘤]