摘要
目的观察阿帕替尼(Apatinib,Apa)和5-氟尿嘧啶(5-Fluorouracil,5-Fu)分别在单药、联合情况下对肠癌细胞系HT-29的增殖抑制及促凋亡作用,并研究其作用机制是否与信号通路磷脂酰肌醇3-激酶(phospholipid inositol 3-kinase,PI3K)/蛋白激酶B(protein kinase B,Akt)相关。方法将细胞分为对照组(加入二甲基亚砜,dimethyl sulfoxide,DMSO)、阿帕替尼组(80μmol·L^-1 Apa)、5-Fu组(100μmol·L^-15-Fu),联合组(80μmol·L^-1 Apa+100μmol·L^-15-Fu)。给药48 h后,用四甲基偶氮唑蓝(Methyl Thiazolyl Tetrazolium,MTT)法检测不同浓度Apa及5-Fu对细胞增殖的影响;用流式细胞术检测Apa及5-Fu对细胞凋亡的影响;用克隆形成实验检测Apa及5-Fu对细胞克隆形成能力的影响;用蛋白质印迹法(Western blot)检测Apa及5-Fu对PI3K、磷酸化磷脂酰肌醇3-激酶(phosphorylated phospholipid inositol 3-kinase,p-PI3K)、Akt、磷酸化蛋白激酶B(phosphorylated protein kinase B,p-Akt)蛋白表达的影响。结果Apatinib 80μmol·L^-1+5-Fu 100μmol·L^-1联合组细胞增殖抑制率提高最显著,此时对照组、阿帕替尼组、5-Fu组和联合组细胞增殖抑制率分别为0%,(0.25±0.08)%,(0.40±0.04)%,(0.74±0.02)%;细胞凋亡率分别为(3.00±1.20)%,(41.60±2.15)%,(43.90±1.80)%,(56.10±1.50)%;细胞克隆数分别为(88.33±3.06),(61.71±1.72),(57.75±4.21),(36.12±2.16)个。阿帕替尼组、5-Fu组、联合组的上述指标与对照组比较,差异均有统计学意义(P<0.05),联合组的上述指标与阿帕替尼组、5-Fu组比较,差异均有统计学意义(P<0.05)。对照组、阿帕替尼组、5-Fu组、联合组细胞p-Akt蛋白表达量分别为1.05±0.10,0.10±0.01,0.80±0.08,0.11±0.01;p-PI3K蛋白表达量分别为0.92±0.06,0.61±0.10,1.01±0.11,0.80±0.05。阿帕替尼组、联合组的上述指标与对照组、5-Fu组比较,差异均有统计学意义(均P<0.05)。结论Apa及5-Fu单药及联合均能以浓度依赖的方式抑制肠癌HT-29细胞系增殖。Apa可通过抑制PI3K/Akt�
Objective To observe the effect of apatinib(Apa)and 5-Fluorouracil(5-Fu)on the proliferation inhibition and apoptosis of HT-29 cell line under the condition of single drug and combination,and to study whether the mechanism is related to phospholipid inositol 3-kinase(PI3K)/protein kinase B(Akt)signal pathway.Methods The HT-29 cells were assigned to control group(dimethyl sulfoxide,DMSO),Apa group(80μmol·L^-1 Apa),5-Fu group(100μmol·L^-1 5-Fu),combination group(80μmol·L^-1 Apa+100μmol·L^-1 5-Fu).After 48 h-treatment,the effects of Apa and 5-Fu at different concentrations on cell proliferation were detected by Methyl Thiazolyl Tetrazolium(MTT)method;the effects of Apa and 5-Fu on cell apoptosis were detected by flow cytometry;the effects of Apa and 5-Fu on cell clonogenesis were detected by clonogenesis experiment;the effects of Apa and 5-Fu on protein expression of PI3K,phosphorylated phospholipid inositol 3-kinase(p-PI3K)、Akt、phosphorylated protein kinase B(p-Akt)were detected by Western blot.Results The cell proliferation inhibition rate of apatinib 80μmol L^-1+5-Fu 100μmol L^-1 combination group was the most significant.At this time,the cell proliferation inhibition rates of control group,apatinib group,5-Fu group and combination group were 0%,(0.25±0.08)%,(0.40±0.04)%,(0.74±0.02)%,cell apoptosis rates were(3.00±1.20)%,(41.60±2.15)%,(43.90±1.80)%,(56.10±1.50)%.The number of cell clones was 88.33±3.06,61.71±1.72,57.75±4.21,36.12±2.16,respectively.The above indexes of apatinib group,5-Fu group and combination group were significantly different from those of the control group(P<0.05),the above indexes of the combined group were statistically significant compared with those of apatinib group and 5-Fu group(P<0.05).P-Akt protein expression was 1.05±0.10,0.10±0.01,0.80±0.08,0.11±0.01 in the control group,apatinib group,5-Fu group and combination group,respectively;p-PI3K protein expression was 0.92±0.06,0.61±0.10,1.01±0.11,0.80±0.05.Compared with the control group and 5-Fu group,the a
作者
张婧超
郑伟
丛秀峰
陈俊
ZHANG Jing-chao;ZHENG Wei;CONG Xiu-feng;CHEN Jun(Department of Oncology,Shengjing Hospital,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
2020年第20期3264-3267,共4页
The Chinese Journal of Clinical Pharmacology
