片仔癀通过BCL-2及Caspase途径参与急性淋巴白血病耐药细胞株MOLT-4/ADM耐药逆转机制的研究  

Pien Tze Huang Participated in the Drug Resistance Mechanism of MOLT-4/ADM Cell Line in Acute Lymphoblastic Leukemia through BCL-2 and Caspase Pathway

作　　者：梅序桥 吴阿阳 刘惠美 黄家福[2] 薛钰[2] 许煜煌 张琳[3] 陆璇 王国伟 胡佳伟 潘裕添[2] MEI Xuqiao;WU Ayang;LIU Huimei;HUANG Jiafu;XUE Yu;XU Yuhuang;ZHANG Lin;LU Xuan;WANG Guowei;HU Jiawei;PAN Yutian(Department of Clinical Laboratory,Zhangzhou Affiliated Hospital of Fujian Medical University,Zhangzhou 363000,China;The Engineering Technological Center of Mushroom Industry,Minnan Normal University,Zhangzhou 363000,China;The Second Ward of Obstetrics,Renmin Hospital of Wuhan University,Wuhan 430000,China;Zhangzhou Skin Disease Prevention and Treatment Hospital of Fujian Province,Zhangzhou 363000,China;Department of Clinical Laboratory,Chaoyang Hospital of Zhangzhou City,Zhangzhou 363000,China)

机构地区：[1]福建医科大学附属漳州市医院检验科,漳州363000 [2]闽南师范大学菌物产业工程技术中心,漳州363000 [3]武汉大学人民医院产科二病区,武汉430000 [4]福建省漳州市皮肤病防治院,漳州363000 [5]福建省漳州市朝阳医院检验科,漳州363000

出　　处：《中国医药指南》2024年第27期71-75,共5页Guide of China Medicine

基　　金：福建省中医药科技项目(2017FJZYZY209);福建省自然科学基金项目(2020J011302);漳州市自然科学基金(ZZ2021J12)。

摘　　要：目的探讨片仔癀对耐阿霉素的急性淋巴细胞白血病细胞株MOLT-4/ADM药物耐受逆转的机制。方法在不同浓度片仔癀作用MOLT-4/ADM细胞48 h后,用MTS法检测MOLT-4/ADM增殖的影响;利用流式细胞术检测MOLT-4/ADM细胞凋亡率;实时荧光PCR检测耐药基因MDR1以及BCL-2 mRNA、Caspase-3/8/9 mRNA表达;蛋白免疫印迹法(western blotting)检测凋亡蛋白BCL-2、Caspase-3/8/9以及多药耐药蛋白(P-gp)表达。结果0.4 mg/ml片仔癀组对MOLT-4/ADM耐阿霉素的IC50逆转倍数为4.02倍。0.2 mg/ml片仔癀组凋亡率为(9.87±0.47)%,0.4 mg/ml片仔癀组凋亡率为(15.17±0.69)%,0.8 mg/ml片仔癀组凋亡率为(20.32±0.87)%,均高于阴性对照组(6.06±0.35)%。随着片仔癀用药浓度的增大,BCL-2 mRNA表达下降,而Caspase-3/8/9 mRNA升高。BCL-2蛋白表达下降,而Caspase-3/9蛋白表达量升高,Caspase-8蛋白相对变化不大。结论片仔癀可以通过BCL-2以及Caspase途径诱导凋亡,增加MOLT-4/ADM对阿霉素的敏感性。Objective To explore the mechanism of Pien Tze Huang on drug reversal of adriamycin resistant human acute lymphoblastic leukemia cell line MOLT-4/ADM.Methods After 48 hours of treatment with different concentrations of Pien Tze Huang,the proliferation of MOLT-4/ADM cells was detected by MTS.The rate of apoptosis in MOLT-4/ADM cells was determined using flow cytometry.qPCR was used to detect the expression of MDR1,BCL-2 and Caspase-3/8/9 mRNA.The expressions of apoptotic protein BCL-2,Caspase-3/8/9 and multidrug resistant protein(P-gp)were detected by Western blotting.Results The IC50 reversal ratio of adriamycin resistant to MOLT-4/ADM in 0.4 mg/ml Pien Tze Huang group was 4.02 times.the apoptosis rate of 0.2 mg/ml Pien Tze Huang group was(9.87±0.47)%,the apoptosis rate of 0.4 mg/ml Pien Tze Huang group was(15.17±0.69)%,the apoptosis rate of 0.8 mg/ml Pien Tze Huang group was(20.32±0.87)%,three different concentration groups of Pien Tze Huang were higher than negative control group(6.06±0.35)%.qPCR showed that with the increase of medication concentration in Pien Tze Huang,the expression of BCL-2 mRNA decreased,while Caspase-3/8/9 mRNA increased.Protein immunoblots showed that with the increase of Pien Tze Huang concentration,the expression of BCL-2 protein decreased,while the expression of Caspase-3/9 protein increased,but not in Caspase-8 protein.Conclusions Pien Tze Huang induce apoptosis through BCL-2 and caspase pathway and increase the sensitivity of MOLT-4/ADM to adriamycin.

关 键 词：片仔癀 急性淋巴细胞白血病 多药耐药 凋亡 

分 类 号：R733.71[医药卫生—肿瘤]