麦门冬汤加减联合顺铂对A549细胞化疗增敏的作用机制  

Mechanism of Modified Maimendong Tang Combined with Cisplatin in Enhancing Chemosensitivity on Human Lung Adenocarcinoma A549 Cells

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作  者:方芳 谢伶俐 宁雪坚 范先基 吴万垠 FANG Fang;XIE Ling-li;NING Xue-jian;FAN Xian-ji;WU Wan-yin(Second Clinical College,Guangzhou University of Chinese Medicine,Guangzhou 510006,China;Guangxi Zhuang Autonomous Region Liuzhou Hospital of Traditional Chinese Medicine ( TCM) ,Liuzhou 545000,China;Guangdong Province Hospital of TCM,Guangzhou 510370,China)

机构地区:[1]广州中医药大学第二临床医学院,广州510006 [2]广西壮族自治区柳州市中医医院,广西柳州545000 [3]广东省中医院,广州510370

出  处:《中国实验方剂学杂志》2019年第4期49-56,共8页China Journal of Experimental Traditional Medical Formulae

基  金:广西壮族自治区卫生厅中医药科技专项( GZPT13-49);广西中医药大学自然科学研究课题项目( 2017QN030).

摘  要:目的:通过体外实验观察麦门冬汤加减(modified Maimendong Tang)联合顺铂(cisplatin)对人肺腺癌A549细胞增殖,凋亡及侵袭转移能力及对半胱氨酸蛋白酶-3(Caspase-3),表皮生长因子受体(epidermal growth factor receptor,EGFR)的蛋白表达量的影响,探讨麦门冬汤加减方化疗增敏的作用并研究其相关机制。方法:分别用麦门冬汤加减(15 g·L-1),顺铂(9 mg·L-1),联合药物干预肺腺癌A549细胞,实验分为空白组、麦门冬汤加减组、顺铂组、麦门冬汤加减+顺铂组。利用噻唑蓝(MTT)比色法检测不同质量浓度(0,5,10,15,20,25 g·L-1)麦门冬汤加减和不同质量浓度(0,3,6,9,12,15 mg·L-1)顺铂干预A549细胞24,48,72 h后的增殖能力,检测各组A549细胞的增殖能力;流式细胞术分析各组的凋亡程度及周期的变化;划痕实验和transwell小室实验检测各组的侵袭转移能力;蛋白免疫印迹法检测各组Caspase-3,EGFR蛋白表达的变化情况。结果:与空白组比较,麦门冬汤加减,顺铂干预A549细胞后呈浓度依赖性、时间依赖性抑制细胞的增殖(P<0.05);各药物组均能抑制A549细胞增殖能力,减弱划痕愈合能力,减少穿过transwell小室的细胞,促进细胞凋亡,下调Caspase-3,EGFR蛋白表达(P<0.05)。与单独给药组比较,麦门冬汤加减与顺铂联合应用后更能明显抑制肺腺癌A549细胞增殖能力和更能诱导A549细胞的凋亡,G0/G1期的细胞增多更明显,S期的细胞明显减少(P<0.05);联合用药后,细胞划痕愈合能力明显减弱,穿过transwell小室的细胞明显减少,Caspase-3,EGFR的蛋白表达量下降更明显(P<0.05)。结论:麦门冬汤加减与顺铂合用后抑制肺癌细胞A549增殖及侵袭转移能力,诱导细胞凋亡,两者存在协同增敏作用,其相关机制可能与下调Caspase-3,EGFR的蛋白表达相关。Objective: To observe effect of modified Maimendong Tang combined with cisplatin on the cell proliferation,apoptosis,invasion and metastasis and the protein expressions of Caspase-3,epidermal growth factor receptor ( EGFR) of human lung adenocarcinoma A549 cells in vitro,so as to investigate their relevant mechanisms in inhibiting cells proliferation,invasion and metastasis and inducing apoptosis of A549 cells. Method: The lung cancer cells A549 were respectively treated with modified Maimendong Tang ( 15 g·L - 1 ) , cisplatin ( 9 mg·L - 1 ) ,and combined drugs. Afterwards,they were divided into control group,modified Maimendong Tang group,cisplatin group and modified Maimendong Tang combined with cisplatin group. Thiazolyl blue tetrazolium bromide ( MTT) assay was used to evaluate the proliferation of A549 cells treated with different concentrations of modified Maimendong Tang ( 0,5,10,15,20,25 g·L - 1 ) and cisplatin ( 0,3,6,9,12, 15 mg·L - 1 ) for 24,48,72 h. The proliferation of A549 cells in each group was detected by MTT assay;flow cytometry was used to detect the degree of apoptosis and cycle in the above four groups of cells;scratch test and transwell migration test were performed to observe the abilities of invasion and metastasis of each group;Western blot was used to detect Caspase-3 and EGFR protein expression. Result: The concentration of modified Maimendong Tang and cisplatin and the time of intervention were negatively correlated with the proliferative capacity of A549 cells ( P < 0. 05);compared with control group,modified Maimendong Tang could reduce the expressions of Caspase-3 and EGFR obviously ( P < 0. 05 );compared with cisplatin group and modified Maimendong Tang group,the modified Maimendong Tang combined with cisplatin group significantly inhibited the proliferation of lung cancer cell and induced its apoptosis,the number of cells in G0 /G1 phase increased apparently,and the cells in S phase decreased significantly ( P < 0. 05);compared with other groups,A549 cells treated with the co

关 键 词:麦门冬汤加减 顺铂 肺癌细胞 细胞凋亡 细胞侵袭转移 

分 类 号:R22[医药卫生—中医基础理论;医药卫生—中医学] R242R2-031R289R285.5

 

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