百里醌通过磷酸化p38MAPK途径介导NSCLC细胞毒性作用的机制研究  

Mechanism of thymoquinone mediates NSCLC cytotoxicity by phosphorylation of p38MAPK pathway

作　　者：陈子盛[1] 廖小雯[1] 张亦飞[1] 肖靖华[1] 陈芸[1] 刘清霞[1] 王鹏[1] 车鹏彪 朱连雨 田东波[1] CHEN Zisheng;LIAO Xiaowen;ZHANG Yifei;XIAO Jinghua;CHEN Yun;LIU Qingxia;WANG Peng;CHE Pengbiao;ZHU Lianyu;TIAN Dongbo(Department of Respiratory Medicine,The Sixth Hospital Affiliated to Guangzhou Medical University,Qingyuan People＇s Hospital,Qingyuan,Guangdong,511500;Department of Neurology Medicine,Wuyi Hospital of Traditional Chinese Medicine,Jiangmen,Guangdong,529000)

机构地区：[1]广州医科大学附属第六医院/清远市人民医院呼吸二病区,广东清远511500 [2]广东省五邑中医院神经内科,广东江门529000

出　　处：《实用临床医药杂志》2018年第11期6-9,14,共5页Journal of Clinical Medicine in Practice

摘　　要：目的探讨百里醌(TQ)对非小细胞肺癌(NSCLC)细胞毒性作用的分子机制。方法 NSCLC鳞癌细胞SK-MES-1接种于96孔板,予20、40、60、80、100μmol/L TQ培养24 h,观察浓度依赖性,计算TQ的IC50值;予接近IC50浓度TQ培养SK-MES-1,观察时间依赖性;予ERK抑制剂U0126和p38抑制剂SB203580分别作用于SK-MES-1、95-D,观察细胞存活率;Western blot检测U0126预处理1 h,加TQ孵育SK-MES-1 30 min后测p-p38、p38,p-ERK1/2、ERK1/2,p-JNK、JNK蛋白表达。结果 (1)TQ呈浓度和时间依赖性降低NSCLC细胞存活率;(2)30μmol/L TQ和10μmol/L U0126+30μmol/L TQ比较其细胞存活率有显著差异(P=0.000),但与10μmol/L SB203580+30μmol/L TQ比较无显著差异(P=1.00),10μmol/L SB203580+30μmol/L TQ与10μmol/L U0126+30μmol/L TQ组比较有显著差异(P=0.000);60μmol/L TQ、10μmol/L U0126+60μmol/L TQ、10μmol/L SB203580+60μmol/L TQ两两比较均无显著差异(P>0.05);随TQ浓度增加,SB203580对95-D细胞的保护作用逐渐减弱,10μmol/L SB203580+40μmol/L TQ组与40μmol/L TQ组比较细胞存活率仍有显著差异(P=0.033);(3)Western blot结果表明U0126显著抑制ERK1/2磷酸化,p38随TQ浓度增加而磷酸化增加,但ERK1/2磷酸化减少,JNK磷酸化无明显改变。结论 TQ透过磷酸化p38途径介导NSCLC毒性作用,而不是ERK1/2途径。Objective To explore the mechanism of thymoquinone（ TQ） on NSCLC cytotoxicity. Methods SK-MES-1 was inoculated into 96-well plates and cultured at 20,40,60,80 and100 μmol/L TQ for 24 h,and the IC50 of TQ was calculated. SK-MES-1 was cultured in close proximity to IC50 concentration TQ,and time-dependent was observed. The ERK inhibitor U0126 and the p38 inhibitor SB203580 were applied to SK-MES-1 and 95-D,respectively,then TQ-activated MAPK-mediated cytotoxicity were observed. The SK-MES-1 expression of p-p38,p38,p-ERK1/2,ERK1/2,pJNK and JNK protein were detected by U0126 pretreatment for 1 h and TQ-cultured for 30 min. Results（1） TQ was used to mediate NSCLC cells in a concentration and time-dependent manner,and the viability of NSCLC cells was decreased.（2） The cell viability of 30 μmol/L TQ and 10 μmol/L U0126 ＋ 30 μmol/L TQ showed significant differences（ P = 0. 000）,but no significant difference was found when compared with 10 μmol/L SB203580 ＋ 30 μmol/L TQ（ P = 1. 00）. 10 μmol/L SB203580 ＋ 30 μmol/L TQ was significantly different from 10 μmol/L U0126 ＋ 30 μmol/L TQ（ P =0. 0 0 0）. 6 0 μmol/L TQ,1 0 μmol/L U0 1 2 6 ＋ 6 0 μmol/L TQ,1 0 μmol/L SB2 0 3 5 8 0 ＋ 6 0 μmol/L TQ showed no significant differences between every two groups（ P 〉 0. 0 5）. With the increase of TQ concentration, the protective effect of SB203580 on 95-D cells gradually decreased, and10 μmol/L SB203580 ＋ 40 μmol/L TQ group was significantly different from 40 μmol/L TQ group（ P = 0. 033）.（3） Western blot analysis showed that U0126 could significantly inhibit the phosphorylation of ERK1/2,phosphorylated p38 increased with the increasing of TQ concentration. However,ERK1/2 phosphorylation decreased,and JNK phosphorylation did not change significantly.Conclusion TQ can mediate NSCLC cytotoxicity through phosphorylation of p38 pathway,but not ERK1/2 pathway.

关 键 词：百里醌 非小细胞肺癌 丝裂原活化蛋白激酶 细胞毒性 

分 类 号：R734.2[医药卫生—肿瘤]