摘要
目的探讨安罗替尼联合5-氟尿嘧啶(5-fluorouracil,5-FU)对结直肠癌细胞增殖和凋亡的作用。方法采用不同浓度的安罗替尼(0、2、10、20、40、80、160和320μmol/L)、5-FU(0、2、10、20、40、80、160和320μmol/L)及安罗替尼(0、2、10、20、40、80、160和320μmol/L)联合5-FU(2μmol/L)作用于结直肠癌SW480和DLD-1细胞,运用Cell Counting Kit-8(CCK-8)检测不同药物浓度对细胞增殖能力的影响。将SW480及DLD-1细胞分为空白对照组、安罗替尼单药组(20μmol/L)以及安罗替尼(20μmol/L)联合5-FU(2μmol/L)组,培养24 h之后进行集落形成实验检测不同处理对细胞的增殖能力的影响。应用流式细胞术检测细胞凋亡情况,采用Western blot法检测细胞凋亡蛋白cleaved-caspase-3表达水平。结果CCK-8结果显示,作用于结直肠癌细胞24 h后,安罗替尼联合5-FU组的半抑制浓度(50%inhibitory concentration,IC50)值(SW480细胞:27.93μmol/L,DLD-1细胞:24.59μmol/L)较安罗替尼单药组(SW480细胞:52.37μmol/L,DLD-1细胞:48.29μmol/L)与5-FU单药组(SW480细胞:163.5μmol/L,DLD-1细胞:152.2μmol/L)降低(均P<0.05),且其对细胞的增殖抑制作用呈浓度依赖性。集落形成实验显示,与对照组比较,安罗替尼单药组SW480和DLD-1细胞集落形成数均下降(均P<0.01)。安罗替尼联合5-FU组SW480和DLD-1细胞集落形成数均较安罗替尼单药组降低(均P<0.01)。流式细胞术显示,安罗替尼单药组的细胞凋亡率[SW480细胞:(20.46±1.06)%,DLD-1细胞:(19.72±0.80)%]较对照组[SW480细胞:(8.13±0.76)%,DLD-1细胞:(7.59±1.21)%]升高(均P<0.01),较安罗替尼联合5-FU组[SW480细胞:(24.14±1.33)%,DLD-1细胞:(36.46±1.05)%]降低(均P<0.05)。安罗替尼单药组cleaved-caspase-3表达水平较对照组均升高(均P<0.01),安罗替尼联合5-FU组cleaved-caspase-3表达水平较安罗替尼单药组均升高(均P<0.01)。结论安罗替尼联合5-FU对人结直肠癌细胞发挥抑制增殖和促进凋亡的作用。
Objective To investigate the effect of anlotinib combined with 5-fluorouracil(5-FU)on the proliferation and apoptosis of colorectal cancer cells.Methods Colorectal cancer cell lines SW480 and DLD-1 were treated with different concentrations of anlotinib(0,2,10,20,40,80,160 and 320μmol/L),5-FU(0,2,10,20,40,80,160 and 320μmol/L),and anlotinib(0,2,10,20,40,80,160 and 320μmol/L)combined with 5-FU(2μmol/L).Cell Counting Kit-8(CCK-8)was used to detect the effects of different concentrations of these drugs on cell proliferation.SW480 and DLD-1 cells were divided into the control group,anlotinib monotherapy group(20μmol/L),and anlotinib(20μmol/L)combined with 5-FU(2μmol/L)group.The colony formation experiment was performed to detect the effects of different treatments on cell proliferation after 24 h culture.Flow cytometry was used to detect the apoptosis of colorectal cancer cells.Western blot was used to detect the expression of cleaved-caspase-3.Results CCK-8 results showed that after treated for 24 h,the 50%inhibitory concentration(IC50)value of the anlotinib combined with 5-FU group(SW480:27.93μmol/L,DLD-1:24.59μmol/L)was lower than that of the anlotinib monotherapy group(SW480:52.37μmol/L,DLD-1:48.29μmol/L)and the 5-FU monotherapy group(SW480:163.5μmol/L,DLD-1:152.2μmol/L;all P<0.05),and the cell proliferation was inhibited in a concentration-dependent manner.Colony formation experiment showed that compared with the control groups,the colony forming numbers of SW480 cells and DLD-1 cells in the anlotinib monotherapy groups both decreased significantly(both P<0.01),and the colony formation numbers of the anlotinib combined with 5-FU groups were both lower than those of the anlotinib monotherapy groups(both P<0.01).Flow cytometry showed that the cell apoptosis rates of the anlotinib monotherapy groups in both cell lines[SW480:(20.46±1.06)%,DLD-1:(19.72±0.80)%]were higher than those of the control groups[SW480:(8.13±0.76)%,DLD-1:(7.59±1.21)%;both P<0.01]and were lower than those of the anlotinib combi
作者
皇甫超男
徐沭芬
丁洁
李娟
汪洋
王科明
Huangfu Chaonan;Xu Shufen;Ding Jie;Li Juan;Wang Yang;Wang Keming(Department of Oncology,the Second Affi liated Hospital of Nanjing Medical University,Nanjing 210011,China)
出处
《实用肿瘤杂志》
CAS
2021年第6期507-513,共7页
Journal of Practical Oncology
基金
国家自然科学基金青年科学基金项目(81802373)
北京医卫健康公益基金会医学科学研究基金资助项目(YWJKJJHKYJJ-B1850242018.1.1~2020.12.31)。
