摘要
目的探究羟基氯喹对人结肠癌SW480细胞增殖及凋亡的影响,并分析可能的机制。方法体外培养人结肠癌细胞系SW480细胞,随机分为空白组和低、中、高剂量实验组。空白组正常培养,低、中、高剂量实验组分别用加入含羟基氯喹终浓度为20,40,80μg·mL^(-1)的细胞培养液。用噻唑蓝(MTT)法检测细胞增殖活性;用平板克隆实验检测细胞克隆形成率;用AnnexinV-FITC/PI双染法检测细胞凋亡情况;用蛋白质印迹法检测Toll样受体4(TLR4)、核转录因子kappa B p65(NF-κB p65)、半胱氨酸的天冬氨酸蛋白酶-3(Caspase-3)蛋白表达水平。结果空白组和低、中、高剂量实验组SW480细胞存活率分别为100%,(83.15±12.45)%,(62.01±9.32)%和(43.25±6.45)%;这4组的克隆形成率分别为(85.16±12.77)%,(65.28±9.78)%,(51.06±7.07)%和(37.11±5.57)%;这4组的凋亡率分别为(9.12±1.37)%,(16.72±2.51)%,(25.89±3.89)%和(39.75±5.97)%;这4组的TLR4蛋白相对表达水平分别为1.15±0.18,0.90±0.13,0.65±0.09和0.29±0.05;这4组的NF-κB p65蛋白相对表达水平分别为1.27±0.20,0.96±0.15,0.69±0.11和0.47±0.08;这4组的Caspase-3蛋白相对表达水平分别为0.43±0.07,0.66±0.09,1.01±0.16和1.38±0.21。上述指标,低、中、高剂量组与空白组比较,差异均有统计学意义(均P<0.05)。结论羟基氯喹可能通过抑制TLR4/NF-κB通路活化抑制人结肠癌SW480细胞增殖并诱导细胞凋亡。
Objective To investigate the effect of hydroxychloroquine on proliferation and apoptosis of human colon cancer SW480 cells and analyze the possible mechanism.Methods Human colon cancer cell line SW480 was cultured in vitro and randomly divided into blank group and experimental-L,-M,-H groups.The blank group was cultured normally;the experimental-L,-M,-H groups were added with hydroxychloroquine,with the final concentration of 20,40 and 80μg·mL^(-1) cell culture medium.Methyl thiazolyl tetrazolium(MTT)assay was used to detect cell proliferation activity;the cell clone formation rate was detected by plate cloning experiment;Annexin V-FITC/PI double staining was used to detect apoptosis;toll like receptor 4(TLR4),nuclear transcription factor kappa B p65(NF-κB p65)and caspase-3 protein expression level were detected by Western blot.Results The survival rates of SW480 cells in the blank group and experimental-L,-M,-H groups were 100%,(83.15±12.45)%,(62.01±9.32)%and(43.25±6.45)%,respectively;the clone formation rates of the five groups were(85.16±12.77)%,(65.28±9.78)%,(51.06±7.07)%and(37.11±5.57)%,respectively;the apoptosis rates of the five groups were(9.12±1.37)%,(16.72±2.51)%,(25.89±3.89)%and(39.75±5.97)%,respectively;the TLR4 proteins of the five groups were 1.15±0.18,0.90±0.13,0.65±0.09 and 0.29±0.05 respectively;the NF-κB p65 of these five groups protein was 1.27±0.20,0.96±0.15,0.69±0.11 and 0.47±0.08 respectively;the caspase-3 proteins of the five groups were 0.43±0.07,0.66±0.09,1.01±0.16 and 1.38±0.21,respectively.For the about indicators,the differences between blank group and experimental-L,-M,-H groups were statistically significant(all P<0.05).Conclusion Hydroxychloroquine may inhibit TLR4/NF-κB Activation of pathway inhibits proliferation and induces apoptosis of human colon cancer SW480 cells.
作者
屈翘
苗超奇
苏琪
QU Qiao;MIAO Chao-qi;SU Qi(General Surgery Department,Shengjing Hospital of China Medical University,Shenyang 110000,Liaoning Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2022年第14期1593-1597,共5页
The Chinese Journal of Clinical Pharmacology
基金
辽宁省科学技术计划基金资助项目(2020JH2)。
关键词
结肠癌
羟基氯喹
增殖
凋亡
TOLL样受体4
human colon cancer sw480 cells
hydroxychloroquine
proliferation
apoptosis
toll-like receptor 4 pathway
