松油烯-4-醇对结直肠癌细胞恶性行为的影响及其机制  

Effects of terpinene-4-ol on malignant behavior of colorectal cancer cells and its mechanism

作　　者：杨艳 吴映敏 曾智锐 李连 张毅 陈腾祥 YANG Yan;WU Ying-min;ZENG Zhi-rui;LI Lian;ZHANG Yi;CHEN Teng-xiang(Dept of Physiology,School of Basic Medicine,Guizhou Medical University,Guiyang 550004,China;Dept of Blood Transfusion,the Third Affiliated Hospital of Guizhou Medical University,Duyun,Guizhou 558000,China;The Affiliated Tumor Hospital of Guizhou Medical University,Guiyang 550004,China)

机构地区：[1]贵州医科大学基础医学院生理学教研室,贵州贵阳550004 [2]贵州医科大学第三附属医院输血科,贵州都匀558000 [3]贵州医科大学附属肿瘤医院,贵州贵阳550004

出　　处：《中国药理学通报》2022年第12期1836-1845,共10页Chinese Pharmacological Bulletin

基　　金：贵州省高层次创新留学人才创新创业项目(留学人才择优资助合同(2018)05号);贵州省教育厅青年科技人才成长项目(黔教合KY字[2018]173号);贵州医科大学高层次人才启动基金项目(校博合J字[2020]066号);贵州医科大学国家自然科学基金培育项目(No 20NSP009)。

摘　　要：目的探究松油烯-4-醇(terpinene-4-alcohol,T4O)对结直肠癌细胞RKO和HCT116恶性行为的影响及机制。方法分别以0、1、2和4μmol·L^(-1)的T4O以及4μmol·L^(-1)的5-Fu处理RKO和HCT116细胞,采用CCK-8、克隆平板、流式细胞术、划痕和Transwell实验检测各组细胞增殖、克隆形成、凋亡、周期、迁移和侵袭情况;采用免疫印迹检测各组细胞中E-cadherin、N-cadherin、p21、CyclinB1和cleaved-Caspase7的表达。基于药效团,分析T4O的作用靶点,探究T4O对作用靶点NR3C1表达及降解速率的影响。构建NR3C1沉默细胞,利用划痕和CCK-8实验探究沉默NR3C1对T4O作用的影响。结果T4O明显抑制RKO和HCT116细胞的增殖、克隆形成、迁移和侵袭,并诱导细胞凋亡和G_(1)期阻滞(P均<0.05),且较同浓度阳性药5-Fu明显;T4O明显减少细胞中N-cadherin和CyclinB1的表达,增加E-cadherin、p21和cleaved-Caspase7的表达(P均<0.05)。药效团分析预测发现T4O有10个作用靶点,其中,抑癌蛋白NR3C1的结合评分最高;T4O处理后,细胞中NR3C1的蛋白表达量明显增加,降解速率明显降低(P均<0.05);抑制NR3C1的表达明显缓解T4O对RKO和HCT116细胞的迁移和增殖抑制(P均<0.05)。结论T4O能够通过维持NR3C1蛋白的稳定性抑制结直肠癌RKO和HCT116细胞的恶性行为。Aim To investigate the effect of terpinene 4-alcohol(T4O)on the malignant behavior of colorectal cancer cell RKO and HCT116 and the underlying mechanism.Methods RKO and HCT116 cells were treated with 0,1,2,4μmol·L^(-1) T4O and 4μmol·L^(-1)5-Fu,respectively.The proliferation,clonal formation,apoptosis,cell cycle,migration and invasion of RKO and HCT116 cells were detected by CCK-8,colony formation,flow cytometry,wound healing and Transwell assay;the expressions of E-cadherin,N-cadherin,p21,CyclinB1 and cleaved-Caspase7 in each group of cells were detected by Western blot.Based on pharmacophore,the target of T4O was analyzed and then the effects of T4O on the expression and degradation rate of NR3C1 were explored.NR3C1 knockdown cells were constructed,and the effects of NR3C1 knockdown on the proliferating and migrating inhibition induced by T4O were detected by wound healing and CCK-8 assay.Results T4O significantly inhibited the proliferation,colony formation,migration and invasion of RKO and HCT116 cells,as well as induced apoptosis and G 1 phase arrest(P all<0.05).The effect of T4O was better than that induced by 5-Fu with the same dose.T4O obviously reduced N-cadherin and Cyclin B1 expression,and elevated the E-cadherin,p21 and cleaved-Caspase7 expression(P all<0.05).A total of 10 targets of T4O were discovered,among which NR3C1 had the highest binding score.After T4O treatment,NR3C1 level in cells increased obviously,and the degradation rate decreased markedly(P<0.05).NRC3C1 knockdown significantly relieved the inhibitory effects of T4O on cell prolfieration and migration(P<0.05).Conclusion T4O can inhibit the malignant behavior of colorectal cancer cells RKO and HCT116 by maintaining the stability of NR3C1 protein.

关 键 词：结直肠癌 松油烯-4-醇 增殖 迁移 侵袭 NR3C1 

分 类 号：R284.1[医药卫生—中药学] R329.28[医药卫生—中医学] R735.35R977.6