氯化两面针碱调节AKT/ERK1/2磷酸化影响肺癌细胞生长、运动、微管形成以及免疫应答  

Nitidine chloride regulates AKT/ERK1/2 phosphorylation and affects growth,motility,microtubule formation and immune response of lung cancer cells

作　　者：张建新[1] 刘雨露 朱子贵 赵红[3] 姚平波 ZHANG Jianxin;LIU Yulu;ZHU Zigui;ZHAO Hong;YAO Pingbo(Department of Thoracic Surgery,Nanhua Hospital Affiliated to Nanhua University,Hengyang 421002,China)

机构地区：[1]南华大学附属南华医院胸外科,衡阳421002 [2]南华大学附属南华医院重症医学科,衡阳421002 [3]南华大学护理学院基础护理教研室,衡阳421001 [4]长沙民政职业技术学院,长沙410004

出　　处：《中国免疫学杂志》2022年第10期1177-1182,共6页Chinese Journal of Immunology

基　　金：湖南省自然科学基金项目(2022JJ60104,2022JJ70039)。

摘　　要：目的:研究氯化两面针碱(NC)对肺癌细胞生长、运动及炎症因子的影响,并阐明其可能机制。方法:以肺癌细胞系A549和NCI-H661为体外研究模型,CCK-8测定不同浓度NC对细胞活力的影响。选择0、3、6、12 nmol/L 4个浓度处理细胞,EdU染色测定细胞增殖,Transwell测定细胞侵袭,微管形成实验测定细胞微管形成能力,RT-PCR检测炎症因子iNOS、TNF-α、IL-4和IL-10 mRNA表达,ELISA检测iNOS、TNF-α、IL-4和IL-10含量,Western blot检测VEGF、N-cadherin和E-cadherin蛋白水平及AKT/ERK1/2通路磷酸化。加入AKT激活剂SC79(4μg/ml)进行验证实验。12周龄雄性BALB/c小鼠作为体外研究对象,检测NC对肿瘤的影响。结果:与对照组比较,当NC浓度>6 nmol/L时,可显著降低细胞活力,减少EdU阳性细胞数量、促进凋亡,抑制细胞侵袭,降低微管形成能力并提高细胞抗炎能力。与NC 0μg组相比,NC可抑制肿瘤生长。结论:NC可抑制肺癌细胞生长、运动,提高其抗炎能力,其机制可能与AKT/ERK1/2通路激活有关。Objective:To study the effects of Nitidine chloride(NC)on lung cancer cell growth,motility and inflammatory factors,and to clarify its possible mechanisms.Methods:The lung cancer cell lines A549 and NCI-H661 were used as research models in vitro.CCK-8 was used to determine the effects of different concentrations of NC on cell viability.Four concentrations of 0,3,6,and 12 nmol/L were selected to treat cells,EdU staining was used to measure cell prolferation,Transwell were used to measure cell invasion,microtubule formation experiment were used to measure the ability of cell microtubule formation,RT-PCR was used to measure inflammatory factors iNOS,TNF-α,IL-4 and IL-10 mRNA expressions,ELISA was used to detect iNOS,TNF-α,IL-4 and IL-10 contents,Western blot was used to detect VEGF,N-cadherin and E-cadherin protein levels and AKT/ERK1/2 pathway phosphorylation.AKT activator SC79(4μg/ml)was added for verification experiment.12-week-old male BALB/c mice were used as the object of in vitro study to detect the effect of NC on tumor.Results:Compared with the control group,when the concentration of NC>6 nmol/L,it could significantly reduced cell viability,reduced the number of EdU-positive cells,promote apoptosis,inhibit the number of cell invasion,reduced the ability of microtubule formation and increased cell anti-inflammatory ability.However,compared with the NC 0μg group,NC could inhibit tumor growth.Conclusion:NC can inhibit the growth and movement of lung cancer cells and improve its anti-inflammatory ability,whose mechanism may be related to the activation of the AKT/ERK1/2 pathway.

关 键 词：氯化两面针碱 肺癌细胞 EdU染色 抗炎 AKT/ERK1/2通路 

分 类 号：R734.2[医药卫生—肿瘤] R329.2[医药卫生—临床医学]