摘要
目的:观察川芎嗪(四甲基吡嗪)对耐药相关ATP结合盒转运体B1(ABCB1)、C1(ABCC1)和G2(ABCG2)表达水平的影响,揭示其逆转人非小细胞肺癌顺铂耐药细胞株A549/DDP耐药的机制。方法:采用细胞增殖(CCK8)实验,分析人肺腺癌细胞株A549及顺铂耐药株A549/DDP对顺铂的耐受性,计算耐药指数。分析A549/DDP对川芎嗪的耐受性,遴选逆转耐药所需的工作浓度并检测逆转效果。设置空白组接种A549细胞,模型组、实验组和阳性对照组均接种A549/DDP细胞;空白组和模型组均用1640完全培养基孵育,实验组用含工作浓度川芎嗪的1640完全培养基孵育,阳性对照组用含Tariquidar的1640完全培养基孵育。提取各组全部蛋白及mRNA,用免疫印迹法(Western Blot)和逆转录-聚合酶链反应(RT-PCR)分析川芎嗪对ABCB1、ABCC1和ABCG2及其m RNA表达水平的影响。结果:A549细胞对顺铂的半数抑制浓度(IC50)为(28.33±3.19)μM,A549/DDP对顺铂的IC50为(983.43±101.67)μM,A549/DDP的耐药指数为34.71。当川芎嗪浓度不高于150μM时,A549/DDP细胞活性无影响,遴选100μM为川芎嗪逆转耐药的工作浓度,1μM作为阳性对照药Tariquidar的工作浓度。川芎嗪逆转耐药指数为7.68,Tariquidar为5.74。Western Blot实验发现,模型组ABCB1、ABCC1和ABCG2的表达水平均高于空白组,在实验组和阳性对照组中均不同程度下调,差异有统计学意义(P<0.05)。RT-PCT实验显示,实验组和阳性对照组ABCB1和ABCC1 mRNA的表达水平与模型组比较均有不同程度的下调(P<0.05),但ABCG2 mRNA的表达水未见明显变化(P>0.05)。结论:川芎嗪可下调ABCB1、ABCC1和ABCG2的表达水平,此可能为逆转A549/DDP细胞对顺铂耐受性的机制之一。
Objective:To study the effects of tetramethylpyrazine on the expression levels of ABCB1,ABCC1 and ABCG2,and to reveal the mechanism of tetramethylpyrazine in reversing human non-small cell lung cancer drug-resistant cell line A549/DDP resistance.Methods:Using cell proliferation experiment(CCK8),the tolerance of human lung adenocarcinoma cell line A549 and its cisplatin-resistant strain A549/DDP to cisplatin was analyzed and the resistance index was calculated.The tolerance of A549/DDP to tetramethylpyrazine was analyzed,the working concentration required to reverse resistance was selected and the reversal effect was detected.The blank group was inoculated with A549 cells,and the model group,experimental group and positive control group were inoculated with A549/DDP cells.The blank group and model group were incubated with 1640 complete medium,the experimental group was incubated with 1640 complete medium containing working concentration of tetramethylpyrazine,and the positive control group was incubated with 1640 complete medium containing Tariquidar.The protein and m RNA of all groups were extracted,and the effects of tetramethylpyrazine on the expression levels of ABCB1,ABCC1 and ABCG2 and their mRNA were analyzed by Western Blot and RT-PCR.Results:A549 cell to cisplatin IC50was(28.33±3.19)μM,A549/DDP-cisplatin IC50was(983.43±101.67)μM,and A549/DDP resistance index was 34.71.When the concentration of tetramethylpyrazine was not higher than 150μM,the activity of A549/DDP cells would not be affected.The concentration of 100μM was selected as the working concentration of tetramethylpyrazine to reverse resistance,and 1μM as the working concentration of the positive control drug Tariquidar.Tetramethylpyrazine reversal resistance index was 7.68and Tariquidar’s was 5.74.Western blotting showed that the expression levels of ABCB1,ABCC1 and ABCG2 in the model group were higher than those in the blank group,and their expression levels were down-regulated in the experimental group and the positive control group t
作者
王小梅
薛春苗
王艳梅
梁玲
张寒蕾
梁艳
WANG Xiaomei;XUE Chunmiao;WANG Yanmei;LIANG Ling;ZHANG Hanlei;LIANG Yan(Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China)
出处
《山东中医药大学学报》
2023年第1期64-70,共7页
Journal of Shandong University of Traditional Chinese Medicine
基金
北京中医药大学青年教师项目(编号:2019-JYB-JS-035)。
关键词
非小细胞肺癌
化疗耐药
川芎嗪
逆转耐药
顺铂
ATP结合盒转运体
non-small cell lung cancer
chemotherapy resistance
tetramethylpyrazine
reversal of drug resistance
cisplatin
ATP-binding cassette transporter(ABC)