氯喹联合顺铂对结肠癌HCT116细胞的增殖和凋亡的影响  

Effects of chloroquine combined with cisplatin on proliferation and apoptosis of colon cancer HCT116 cells

作　　者：周静[1] 张配 葛少波[1] 刘婕[1] 张杰[1] ZHOU Jing;ZHANG Pei;GE Shao-bo;LIU Jie;ZHANG Jie(Department of Pharmacy,The First People’s Hospital of Bengbu,Bengbu 233000,Auhui Province,China;School of Pharmacy,Bengbu Medical College,Bengbu 233030,Auhui Province,China)

机构地区：[1]蚌埠市第一人民医院药剂科,安徽蚌埠233000 [2]蚌埠医学院药学院,安徽蚌埠233030

出　　处：《中国临床药理学杂志》2022年第3期224-227,242,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81603155);安徽省自然科学基金青年基金资助项目(1708085QH212)。

摘　　要：目的探讨氯喹(CQ)联合顺铂(DDP)对人结肠癌HCT116细胞增殖和凋亡的影响及其可能机制。方法将HCT116细胞分组为16组:空白对照组(不加药物)、5个浓度(5,10,20,40,80μmol·L^(-1))氯喹-1组至氯喹-5组,5个浓度(2,4,8,16,32μmol·L^(-1))顺铂-1组至顺铂-5组和5个浓度(20μmol·L^(-1)氯喹+2,4,8,16,32μmol·L^(-1)顺铂)联合-1组至联合-5组,各组均培养24 h。用噻唑蓝法检测细胞的存活率,用PI单染流式细胞术检测细胞的凋亡情况,用蛋白质印迹法检测B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)等凋亡相关蛋白的表达水平(灰度值)。结果空白对照组、氯喹-1组至氯喹-5组、顺铂-1组至顺铂-5组和联合-1组至联合-5组于24 h的细胞存活率分别为(100.00±1.05)%,(99.65±1.98)%,(97.43±0.72)%,(83.88±0.51)%,(69.65±3.33)%,(51.87±0.49)%,(98.55±1.50)%,(92.33±0.50)%,(81.66±4.29)%,(70.41±0.50)%,(48.34±1.50)%,(78.61±4.37)%,(59.01±3.52)%,(37.23±0.57)%,(32.88±3.96)%和(21.09±2.57)%。空白对照组、氯喹-3组、顺铂-3组和联合-3组的细胞凋亡率分别为(1.63%±0.50%),(15.97%±1.40%),(24.83%±2.12%)和(47.23±4.29)%;这4组的Bcl-2相对表达量分别为1.11±0.01,0.92±0.03和0.73±0.08,0.46±0.07;这4组的Bax蛋白相对表达量分别为0.18±0.01,0.27±0.03,0.46±0.09和0.58±0.04。氯喹联合顺铂能显著减少抗凋亡Bcl-2蛋白的表达,并上调促凋亡Bax蛋白的表达。联合组与单用组比较,上述指标的差异均有统计学意义(均P<0.05);各给药组与空白对照组比较,上述指标的差异均有统计学意义(均P<0.05)。结论氯喹可以增强顺铂对HCT116细胞的增殖抑制作用,同时可以增强顺铂诱导结肠癌细胞的凋亡,其机制可能与下调Bcl-2和上调Bax有关联。Objective To investigate the effects of chloroquine(CQ)combined with cisplatin(DDP)on proliferation and apoptosis of human colon cancer HCT116 cells,and its possible mechanism.Methods HCT116 cells were divided into 16 groups:blank control group(without drugs),five concentrations(5,10,20,40 and 80μmol·L^(-1))of CQ-1,CQ-2,CQ-3,CQ-4,CQ-5 groups,five concentrations(2,4,8,16,32μmol·L^(-1))of DDP-1,DDP-2,DDP-3,DDP-4,DDP-5 groups,and five concentrations(20μmol·L^(-1) CQ combined with 2,4,8,16,32μmol·L^(-1) DDP)of combined-1,c ombined-2,combined-3,combined-4,combined-5 groups,for 24 h.Cell viability was detected by methyl thiazolyl tetrazolium.The apoptosis of cells was assessed using flow cytometry with PI staining.The relative expressions(gray value)of B-cell lymphoma-2(Bcl-2)and Bcl-2-associated X(Bax)were analyzed using Western blotting.Results The cell survival rate at 24 h in blank control group,CQ-1,CQ-2,CQ-3,CQ-4,CQ-5 groups,DDP-1,DDP-2,DDP-3,DDP-4,DDP-5 groups,combined-1,combined-2,combined-3,combined-4,combined-5 groups were(100.00±1.05)%,(99.65±1.98)%,(97.43±0.72)%,(83.88±0.51)%,(69.65±3.33)%,(51.87±0.49)%,(98.55±1.50)%,(92.33±0.50)%,(81.66±4.29)%,(70.41±0.50)%,(48.34±1.50)%,(78.61±4.37)%,(59.01±3.52)%,(37.23±0.57)%,(32.88±3.96)%,and(21.09±2.57)%.The apoptosis rate in balnk control group,CQ-3 group,DDP-3 group and combined group were(1.63±0.50)%,(15.97±1.40)%,(24.83±2.12)%,(47.23±4.29)%;the relative expressions of Bcl-2 protein in the 4 groups were 1.11±0.01,0.92±0.03,0.73±0.08,0.46±0.07;the relative expressions of Bax protein in the 4 groups were 0.18±0.01,0.27±0.03,0.46±0.09,0.58±0.04.Both combined group could significantly down regulate the expression of anti apoptotic Bcl-2 protein and up regulate the expression of Pro apoptotic Bax protein.Comparison between combined group and CQ group,DDP group,the difference of the factors were significant(all P<0.05);compared between each treatment groups and blank control group,the difference of the factors were significant(all P<0.

关 键 词：结肠癌 氯喹 顺铂 增殖抑制 凋亡 

分 类 号：R97[医药卫生—药品]