去氧地胆草素对肺癌细胞自噬和mTOR信号通路的影响  

Effects of deoxydebilirubin on autophagy and mTOR signaling pathway in lung cancer cells

作　　者：杨钊 马凯 王壮壮 周进才 YANG Zhao;MA Kai;WANG Zhuangzhuang;ZHOU Jincai(Department of Thoracic Surgery,Xi'an International Medical Center Hospital,Xi'an 710100,China)

机构地区：[1]西安国际医学中心医院胸腔外科,陕西西安710100

出　　处：《西部医学》2024年第12期1741-1747,共7页Medical Journal of West China

基　　金：陕西省重点研发计划项目(2022SF-325)。

摘　　要：目的 探究去氧地胆草素(DET)对肺癌细胞自噬和哺乳动物雷帕霉素靶蛋白(mTOR)信号通路的影响。方法 用不同浓度的DET(0、5、10、20和50μM)处理人非小细胞肺癌细胞系(A549)后,使用四甲基偶氮唑盐(MTT)法检测细胞活力,流式细胞仪检测细胞凋亡,Transwell检测细胞迁移和侵袭。通过Western blot检测细胞中Beclin-1、微管相关蛋白1轻链3(LC3)、mTOR、p-mTOR、p70核糖体S6蛋白激酶(P70S6K)、p-P70S6K、真核细胞翻译起始因子4E-结合蛋白1(4E-BP1)和p-4E-BP1的蛋白表达。将12只A549细胞移植瘤裸鼠随机分为对照组和DET组,每组6只。DET组小鼠腹腔注射DET溶液(10 mg/kg),对照组小鼠腹腔注射含1%DMSO的生理盐水,给药21 d后分别检测两组裸鼠的肿瘤体积和重量。通过免疫组化检测肿瘤组织中p-mTOR、p-p70S6K和p-4E-BP1的蛋白表达。结果 与0μM比较,5、10、20、50μM DET处理的A549细胞的生长抑制率升高(P<0.05),凋亡率升高(P<0.05),迁移和侵袭细胞数量降低(P<0.05),Beclin-1和LC3-Ⅱ/LC3-Ⅰ的蛋白相对表达量升高(P<0.05),p-mTOR/mTOR、p-p70S6K/p70S6K和p-4E-BP1/4E-BP1的蛋白相对表达量降低(P<0.05)。与对照组比较,DET组小鼠的肿瘤体积和重量均减小(P<0.05),肿瘤组织中p-mTOR、p-p70S6K和p-4E-BP1的染色评分均降低(P<0.05)。结论 DET的抗肺癌作用可能与其抑制mTOR信号通路并诱导肺癌细胞自噬有关。Objective To reveal the effect of deoxydebilirubin(DET)on autophagy and mammalian target of rapamycin(mTOR)signaling pathway in lung cancer cells.Methods After treatment of human non-small cell lung cancer cell line(A549)with different concentrations of DET(0μM,5μM,10μM,20μM and 50μM),cell viability was detected by tetramethylazolium salt(MTT),apoptosis was detected by flow cytometry,and cell migration and invasion were detected by Transwell.The protein expressions of Beclin-1,microtubule associated protein 1 light chain 3(LC3),mTOR,p-mTOR,p70 ribosomal S6 protein kinase(P70S6K),p-P70S6K,eukaryotic translation initiation factor 4E-binding protein 1(4E-BP1)and p-4E-BP1 were detected by Western blot.Nude mice transplanted with A549 cells were randomly divided into Control group(n=6)and DET group(n=6).The mice in DET group were intraperitoneally injected with DET solution(10 mg/kg),and the mice in Control group were injected intraperitoneally with normal saline containing 1%DMSO.After 21 days of administration,the tumor volume and weight of the two groups of nude mice were detected respectively.The protein expressions of p-mTOR,p-p70S6K and p-4E-BP1 in tumor tissues were detected by immunohistochemistry.Results Compared with 0μM,the growth inhibition rate and apoptosis rate of A549 cells treated with 5μM,10μM,20μM and 50μM DET increased,the number of migratory and invasive cells decreased(P<0.05),the relative expression of Beclin-1 and LC3-Ⅱ/LC3-Ⅰincreased,the relative expression of p-mTOR/mTOR,p-p70S6K/p70S6K and p-4E-BP1/4E-BP1 decreased(P<0.05).Compared with Control,the volume and weight of tumor in DET group decreased,and the staining scores of p-mTOR,p-p70S6K and p-4E-BP1 in tumor tissue decreased(P<0.05).Conclusion The anti-lung cancer effect of DET may be related to its inhibition of mTOR signaling pathway and induction of autophagy in lung cancer cells.

关 键 词：肺癌 去氧地胆草素 自噬 MTOR信号通路 天然药物 

分 类 号：R734.2[医药卫生—肿瘤]