鸦胆子油乳剂对食管鳞状细胞癌TE-1细胞增殖、凋亡和自噬的影响及其可能的机制  

Effects of brucea javanica oil emulsion on proliferation,apoptosis and autophagy of esophageal squamous cell carcinoma TE-1 cells and its possible mechanism

作　　者：付皓云[1] 李嫚[1] FU Haoyun;LI Man(Department of Integrated Traditional Chinese and Western Medicine,Hubei Cancer Hospital,Wuhan 430079,Hubei,China)

机构地区：[1]湖北省肿瘤医院中西医结合科,湖北武汉430079

出　　处：《中国肿瘤生物治疗杂志》2023年第7期560-567,共8页Chinese Journal of Cancer Biotherapy

基　　金：国家中医药管理局第七批全国名老中医药专家学术继承项目[No.国中医药人教函(2022)76号]。

摘　　要：目的:探讨鸦胆子油乳剂(BJOE)对食管鳞状细胞癌TE-1细胞增殖、凋亡和自噬的影响及其可能的机制。方法:按干预措施的不同,将TE-1细胞分为对照组、RAPA(自噬激动剂)组、740Y-P(PI3K激活剂)组、BJOE组、BJOE+RAPA组和BJOE+740Y-P组。采用FCM、克隆形成、Transwell实验检测细胞凋亡、增殖、迁移和侵袭能力,qPCR法检测细胞中PI3K、Akt、mTOR、LC3Ⅰ、LC3Ⅱ、p62、Beclin 1、caspase-3的mRNA表达,WB法检测PI3K、Akt、mTOR及其磷酸化、LC3Ⅱ/Ⅰ、p62、Beclin 1、caspase-3的蛋白表达。结果:与对照组比较,RAPA组和BJOE组细胞凋亡率均显著升高(均P<0.01),细胞克隆形成率、迁移和侵袭能力均显著降低(均P<0.01),细胞中PI3K、Akt、mTOR的mRNA和蛋白磷酸化水平均显著降低(均P<0.05),p62的mRNA和蛋白水平显著降低(均P<0.01),LC3Ⅱ/Ⅰ、Beclin 1和caspase-3的mRNA和蛋白水平显著升高(均P<0.05),740Y-P组的结果则相反(均P<0.05);与RAPA组或740Y-P组比较,BJOE+RAPA组或BJOE+740Y-P组细胞凋亡率显著升高(均P<0.01),克隆形成率、细胞侵袭和迁移能力显著降低(均P<0.01),PI3K、Akt、mTOR的mRNA和蛋白磷酸化水平均显著降低(均P<0.05),p62的mRNA和蛋白水平均显著降低(均P<0.05),LC3Ⅱ/Ⅰ、Beclin 1和caspase-3 mRNA和蛋白水平显著升高(均P<0.05)。结论:BJOE显著抑制TE-1细胞增殖、迁移、侵袭并促进细胞凋亡与自噬,其机制可能与抑制PI3K/Akt/mTOR信号通路的激活有关。Objective:To explore the effects of brucea javanica oil emulsion(BJOE)on proliferation,apoptosis and autophagy of esophageal squamous cell carcinoma TE-1 cells and the possible mechanism.Methods:According to the different interventions TE-1 cells were divided into control group,RAPA(autophagy agonist)group,740Y-P(PI3K activator)group,BJOE group,BJOE+RAPA group and BJOE+740Y-P group.Cell apoptosis,proliferation,migration and invasion were detected by FCM,clonogenesis and Transwell assay;the mRNA expression of PI3K,Akt,mTOR,LC3Ⅰ,LC3Ⅱ,p62,Beclin 1 and caspase-3 in cells was detected by qPCR;and the protein expression levels of PI3K,Akt,mTOR and their phosphorylation as well as the protein expression of LC3Ⅱ/Ⅰ,p62,Beclin1 and caspase-3 were detected by Western blotting.Results:Compared with the control group,RAPA group and BJOE group exhibited increased cell apoptosis rate(both P<0.01),reduced clone formation rate,cell invasion and migration ability(all P<0.01),decreased mRNA and protein phosphorylation levels of PI3K,Akt,mTOR(all P<0.05)as well as decreased mRNA and protein levels of p62(all P<0.01),and increased mRNA and protein levels of LC3Ⅱ/Ⅰ,Beclin 1 and caspase-3(all P<0.05);however,the results of 740Y-P group were opposite(all P<0.05).Compared with RAPA group or 740Y-P group alone,BJOE+RAPA or BJOE+740Y-P group exhibited increased apoptosis rate(P<0.01),decreased clone formation rate,cell invasion and migration ability(all P<0.01),decreased mRNA and protein phosphorylation levels of PI3K,Akt,mTOR(all P<0.05)as well as decreased mRNA and protein levels of p62(all P<0.05),and increased mRNA and protein levels of LC3Ⅱ/Ⅰ,Beclin 1 and caspase-3(all P<0.05).Conclusion:BJOE significantly inhibits the proliferation,invasion,migration and promotes apoptosis and autophagy of TE-1 cells,and the mechanism may be related to inhibition of the activation of PI3K/Akt/mTOR signaling pathway.

关 键 词：鸦胆子油乳剂 食管鳞状细胞癌 TE-1细胞 增殖 凋亡 自噬 PI3K/Akt/mTOR信号通路 

分 类 号：R735.1[医药卫生—肿瘤] R730.2[医药卫生—临床医学]