阿帕替尼联合曲妥珠单抗对胃癌细胞的协同增敏作用

Synergistic sensitization of apatinib and trastuzumab on gastric cancer cells

目的观察阿帕替尼联合曲妥珠单抗杀伤胃癌NCI-N87细胞的协同增敏作用并探讨可能作用机制。方法CCK-8法检测空白对照组、曲妥珠单抗组(0.1、1、10μg/ml)、阿帕替尼组(1μmol/L)及曲妥珠单抗(0.1、1、10μg/ml)+阿帕替尼(1μmol/L)组对NCI-N87细胞的增殖抑制作用,流式细胞术检测NCI-N87细胞凋亡,Western blotting检测HER-2、VEGFR2、Bax、Bcl-2蛋白表达。结果CCK-8检测提示曲妥珠单抗、阿帕替尼能够抑制NCI-N87细胞增殖,在一定浓度范围内作用呈浓度依赖性和时间依赖性(P<0.01);q值计算提示曲妥珠单抗与阿帕替尼具有协同抑制NCI-N87细胞增殖的作用。流式细胞术检测显示联合组NCI-N87胃癌细胞凋亡较单药组明显升高(P<0.05),其中空白对照组、阿帕替尼组(1μmol/L)、曲妥珠单抗(0.1μg/ml)组及曲妥珠单抗(0.1μg/ml)+阿帕替尼(1μmol/L)组的凋亡率分别为(3.0±1.28)%、(5.8±1.63)%、(8.0±3.92)%和(21.6±6.85)%。曲妥珠单抗+阿帕替尼组与空白对照组、曲妥珠单抗及阿帕替尼单药组比较,HER-2蛋白表达显著下调(P<0.05);曲妥珠单抗+阿帕替尼组与空白对照组比较,Bcl-2蛋白表达、Bcl-2/Bax比值明显降低(P<0.01)。结论阿帕替尼联合曲妥珠单抗可能通过下调HER-2蛋白及调控凋亡相关蛋白Bcl-2、Bax的表达,协同抑制NCI-N87细胞增殖和促进细胞凋亡。

Objective To observe the synergistic sensitization effect of apatinib combined with trastuzumab on NCI-N87 cells and its possible mechanism.Methods CCK-8 method was used to detect the inhibitory effect of blank control group,trastuzumab group(0.1,1,10μg/ml),apatinib group(1μmol/L),trastuzumab(0.1,1,10μg/ml)+apatinib(1μmol/L)on the proliferation of NCI-N87 cells.Flow cytometry was used to detect the apoptosis of NCI-N87 cells.Western blotting was used to detect the expression of HER-2,VEGFR2,Bax and Bcl-2 proteins.Results CCK-8 detection indicated that trastuzumab and apatinib could inhibit the proliferation of NCI-N87 cells in a concentration dependent and time-dependent manner(P<0.01);Q-value calculation indicated that trastuzumab and apatinib could synergistically inhibit the proliferation of NCI-N87 cells.Flow cytometry showed that the apoptosis of NCI-N87 gastric cancer cells in the combined group was significantly higher than that in the single drug group(P<0.05).Compared with the blank control group,trastuzumabgroup and apatinib group,the expression of HER-2 protein in trastuzumab+apatinib group decreased significantly(P<0.05);compared with the blank control group,the expression of Bcl-2 protein and the ratio of Bcl-2/Bax in trastuzumab+apatinib group decreased significantly(P<0.01).Conclusion The combination of apatinib and trastuzumab may inhibit the proliferation and promote the apoptosis of NCI-N87 cells by down regulating the expression of HER-2,Bcl-2 and Bax.