山柰酚逆转MCF-7/阿霉素细胞耐药机制研究  

Study on the mechanism of reversal effect of kaempferol on MCF-7/adriamycin cells

作　　者：阿地力江·萨吾提 木塔力甫·艾买提 周文婷 艾尼瓦尔·吾买尔 SAWUTI Adilijiang;MUTALIFU Aimaiti;ZHOU Wen-ting;WUMAIER Ainiwaer(Department of Pharmacology,College of Pharmacy,Urumqi 830011,Xinjiang Uyghur Autonomous Region,China;Central Laboratory,Xinjiang Medical University,Urumqi 830011,Xinjiang Uyghur Autonomous Region,China)

机构地区：[1]新疆医科大学药学院药理教研室,新疆乌鲁木齐830011 [2]新疆医科大学中心实验室,新疆乌鲁木齐830011

出　　处：《中国临床药理学杂志》2021年第14期1806-1810,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81560586);新疆维吾尔自治区自然科学基金资助项目(2016D01C161);新疆维吾尔自治区研究生创新创业启动基金资助项目(XJ2019G193);新疆自治区“十三五”重点学科建设基金资助项目(2016)。

摘　　要：目的研究山柰酚逆转人乳腺癌耐阿霉素细胞MCF-7/阿霉素(ADR)耐药机制。方法(1)将MCF-7和MCF-7/ADR细胞分为空白组(培养基)、对照组(细胞、培养基)和实验组。实验组分别用含不同浓度(20,50,100,150,200,300,400,500μmol·L^(-1))山柰酚的培养基培养,每组分别干预24,48和72 h。干预后MTT法测定细胞存活率。(2)将MCF-7和MCF-7/ADR细胞分为空白组(培养基)、对照组(细胞、培养基)和联合组,联合组分别用含有不同浓度(3,6,12,24,48μmol·L^(-1))阿霉素、不同浓度(3,6,12,24,48μmol·L^(-1))紫杉醇和不同浓度(10,30,60,90,120μmol·L^(-1))顺铂的培养基培养,干预48 h。MTT法测定细胞存活率,计算IC50和耐药倍数。(3)将MCF-7/ADR细胞分为ADR组和低、中2个浓度ADR联用组(ADR+山柰酚20,50μmol·L^(-1));紫杉醇组和低、中2个浓度紫杉醇联用组(紫杉醇+山柰酚20,50μmol·L^(-1));顺铂组和低、中2个浓度顺铂联用组(顺铂+山柰酚20,50μmol·L^(-1)),计算IC50和逆转耐药倍数。(4)细胞分为MCF-7组与MCF-7/ADR组;再将MCF-7/ADR细胞分为4组:空白组和低、中、高3个浓度山柰酚组(山柰酚:20,50,100μmol·L^(-1))。蛋白质印迹法测定核因子κB(NF-κB)、NF-κB抑制剂(I-κB)、多药耐药蛋白(ABCB1)和乳腺癌耐药蛋白(ABCG2)蛋白表达水平(灰度值比值)。结果(1)山柰酚显著抑制MCF-7和MCF-7/ADR细胞存活率,与空白组相比,差异有统计学意义(P<0.05)。(2)MCF-7细胞:ADR组、紫杉醇组和顺铂组的IC50分别为(1.84±0.22),(1.76±0.12)和(0.78±0.53)μmol·L^(-1)。MCF-7/ADR细胞:这3组的IC50分别为(43.68±0.76),(85.93±9.04)和(203.20±5.08)μmol·L^(-1)。2种细胞的IC50相比差异均有统计学意义(均P<0.01)。上述3种药物在MCF-7/ADR细胞的耐药倍数分别为23.66,9.50和257.86。(3)MCF-7/ADR细胞:ADR组和低、中2个浓度联合组的IC50分别为(43.68±0.76),(17.68±3.14)和(23.51±0.06)μmol·L^(-1),低、中2个浓度联合组与ADR组相比差异有统计学意义Objective To investigate the reversal activity and mechanism of kaempferol against MCF-7/adriamycin(ADR)cells.Methods(1)MCF-7 and MCF-7/ADR cells were divided into blank group(containing medium only),control group(not containing drugs)and experimental group.The experimental group was treated with different concentrations of kaempferol(0,20,50,100,150,200,300,400,500μmol·L^(-1))in 24,48,72 h,respectively.After administration,cell viability was measured by MTT assay.(2)Again,the experimental group of MCF-7 and MCF-7/ADR cells were treated with different concentrations of ADR(3,6,12,24,48μmol·L^(-1)),paclitaxel(3,6,12,24,48μmol·L^(-1))and cisplatin(10,30,60,90,120μmol·L^(-1)),after treatment of 48 h,cell viability was measured by MTT assay,and the IC50 and fold of resistance was calculated.(3)MCF-7 cells were divided into 3 groups:adriamycin group,and joint-L,-M groups(adriamycin+20,50μmol·L^(-1)kaempferol);paclitaxel,joint-L,-M groups(paclitaxel+20,50μmol·L^(-1)kaempferol);cisplatin,joint-L,-M groups(cisplatin+20,50μmol·L^(-1)kaempferol).The IC50 and the reverse fold was calculated.(4)The cells were divided into two groups:MCF-7 group and MCF-7/ADR group;the MCF-7/ADR cells were divided into 4 groups:blank group,kaempferol-L,-M,-H groups(kaempferol 20,50,100μmol·L^(-1)),respectively.The expression(ratio of gray value)of nuclear factor-κB(NF-κB),inhibitor of NF-κB(I-κB),multidrug resistance protein(ABCB1),and breast cancer resistance protein(ABCG2)was determined by Western blot.Results(1)Kaempferol significantly inhibited the proliferation of MCF-7 in different time(P<0.01).(2)The IC50 of adriamycin group,paclitaxel group,cisplatin group in MCF-7cells were(1.84±0.22),(1.76±0.12)and(0.78±0.53)μmol·L^(-1),respectively.The IC50 of the 3 groups in MCF-7/ADR cells were(43.68±0.76),(85.93±9.04)and(203.20±5.08)μmol·L^(-1),respectively.There are significant difference between these two groups(P<0.01).The fold of resistance of adriamycin,paclitaxel,cisplatin were 23.66,9.50 and 257.86,respective

关 键 词：山柰酚 乳腺癌 耐药逆转 核因子-ΚB 多药耐药蛋白 阿霉素 

分 类 号：R28[医药卫生—中药学]