阿帕替尼对人乳腺癌细胞株MCF-7细胞生物学行为的影响

Effects of apatinib on biological behavior of human breast cancer cell line MCF-7

目的探讨阿帕替尼对人乳腺癌细胞株MCF-7细胞生物学行为的影响。方法取对数生长期的人乳腺癌MCF-7细胞,随机分为对照组和阿帕替尼组,对照组不予任何处理,阿帕替尼组分别给予浓度为2.5μmol/L、5.0μmol/L、10μmol/L、20μmol/L、40μmol/L的阿帕替尼进行处理。采用二甲基四氮唑蓝(MTT)法检测浓度分别为2.5μmol/L、5.0μmol/L、10μmol/L、20μmol/L、40μmol/L的阿帕替尼对MCF-7细胞生长抑制的作用;采用丫啶橙(AO)/溴乙锭(EB)染色法,于倒置荧光显微镜下观察MCF-7细胞形态变化;采用酶联免疫吸附法测定阿帕替尼对MCF-7细胞上清液血管内皮生长因子(VEGF)含量的影响;采用流式细胞仪检测阿帕替尼对MCF-7细胞周期和凋亡率的影响。结果阿帕替尼对MCF-7细胞具有明显的生长抑制作用,其抑制率随着浓度的增加而升高,呈明显的时间-剂量依赖关系(P<0.05);对照组细胞结构正常,细胞核呈绿色;而阿帕替尼组细胞形态发生明显改变,染色质呈橘红色,呈凋亡状;与对照组细胞周期比较,干预组与对照组G 0/G 1期、S期、G 2/M期均值比率无明显统计学差异(P>0.05);干预组VEGF分泌量明显下降,阿帕替尼浓度越高,VEGF分泌量越低(P<0.05);阿帕替尼组细胞凋亡率明显高于对照组,差异有统计学意义(P<0.05)。结论阿帕替尼可抑制人乳腺癌MCF-7细胞增殖并诱导其凋亡,且具有浓度依赖性,可能与减少VEGF的分泌有关。

Objective To explore the effects of apatinib on the biological behavior of human breast cancer cell line MCF-7.Methods Human breast cancer MCF-7 cells in the logarithmic growth phase were selected and randomly divided into control group and apatinib group.Control group was not given any treatment,and apatinib group was given apatinib at concentrations of 2.5μmol/L,5.0μmol/L,10μmol/L,20μmol/L and 40μmol/L for treatment.MTT assay was used to detect the effects of apatinib at concentrations of 2.5μmol/L,5.0μmol/L,10μmol/L,20μmol/L and 40μmol/L on the growth inhibition of MCF-7 cells.The morphology of MCF-7 cells was observed under inverted fluorescence microscope by acridine orange(AO)/ethidium bromide(EB)staining.The effects of apapitatin on the vascular endothelial growth factor(VEGF)content in supernatant of MCF-7 cells was determined by enzyme-linked immunosorbent assay(ELISA).Flow cytometry was used to detect the effects of apatinib on cell cycle and apoptosis rate of MCF-7.Results Apatinib had obvious growth inhibitory effect on MCF-7 cells,and its inhibition rate was increased with the increase of concentration,showing a time-dose-dependent relationship(P<0.05).The cell structure in control group was normal with green cell nucleus.The cell morphology in apatinib group was obviously changed with orange-red and apoptotic chromatin.Compared with cell cycle in control group,there were no significant differences in the mean value proportion of G 0/G 1 phase,S phase and G 2/M phase between intervention group and control group.(P>0.05).The secretion of VEGF was decreased significantly in intervention group.The higher the concentration of apatinib,the lower the secretion of VEGF(P<0.05).The apoptosis rate in apatinib group was significantly higher than that in control group(P<0.05).Conclusion Apatinib can inhibit the proliferation and induce apoptosis of human breast cancer MCF-7 cells,and it has concentration-dependence which may be related to the decrease of VEGF secretion.