基于VEGF/PI3K/AKT通路探讨消岩汤联合阿帕替尼抑制胃癌MGC-803细胞增殖的作用机制研究

Exploring the mechanism of Xiaoyan Decoction combined with apatinib in inhibiting the proliferation of gastric cancer MGC-803 cells based on the VEGF/PI3K/AKT pathway

[目的]明确消岩汤通过血管内皮生长因子(VEGF)/磷脂酰肌醇3激酶(PI3K)/丝苏氨酸蛋白激酶(AKT)通路治疗胃癌的作用机制。[方法]通过CCK-8检测消岩汤对MGC-803细胞增殖的影响;通过流式细胞术检测消岩汤对MGC-803细胞凋亡和周期的影响;通过蛋白免疫印迹(Western blot)检测消岩汤、阿帕替尼及其联合用药作用于MGC-803细胞后VEGF/PI3K/AKT信号通路相关蛋白的表达。[结果]消岩汤呈剂量依赖性抑制MGC-803细胞的增殖(P<0.05),其半抑制浓度(IC_(50))为63.56 mg/mL;随着消岩汤浓度的增加,MGC-803细胞的总凋亡率逐渐升高(P<0.05);随着消岩汤浓度的增加,MGC-803细胞的G0/G1期细胞所占百分比逐渐降低(P<0.05),S期细胞所占百分比逐渐升高(P<0.05),G2/M期细胞所占百分比变化不大(P>0.05);与空白对照组比较,消岩汤组、阿帕替尼组及其联合用药组的p-PI3K、p-AKT、Bcl-2、血管内皮生长因子A(VEGFA)和血管内皮细胞生长因子受体2(VEGFR-2)蛋白的表达量显著下调(P<0.05),Cleaved Caspase-9和Bax蛋白的表达量显著上调(P<0.05),相较于单药组,联合用药组对相关蛋白的调控作用更强。[结论]消岩汤能有效抑制人胃癌MGC-803细胞增殖,诱导凋亡以及S期阻滞,抗胃癌作用机制与其对VEGF/PI3K/AKT信号通路的调控相关。

[Objective]To investigate the antitumor mechanism of Xiaoyan Decoction on human gastric cancer MGC-803 cells.[Methods]The effect of Xiaoyan Decoction on the proliferation of MGC-803 cells was detected by CCK-8;The effect of Xiaoyan Decoction on apoptosis and cell cycle of MGC-803 cells was detected by flow cytometry;Western blot was used to detect the expression of VEGF/PI3K/Akt signal pathway related proteins after Xiaoyan Decoction,apatinib and their combination on MGC-803 cells.[Results]Xiaoyan Decoction inhibited the proliferation of MGC-803 cells in a dose-dependent manner(P<0.05),and its IC_(50)was 63.56 mg/mL;With the increase of Xiaoyan Decoction concentration,the total apoptosis rate of MGC-803 cells increased gradually(P<0.05);With the increase of Xiaoyan Decoction concentration,the percentage of cells in G0/G1 phase of MGC-803 cells decreased gradually(P<0.05),the percentage of cells in S phase increased gradually(P<0.05),and the percentage of cells in G2/M phase changed little(P>0.05);Compared with the blank control group,the expressions of p-PI3K,p-AKT,Bcl-2,VEGFA and VEGFR-2 proteins in Xiaoyan Decoction group,apatinib group and their combination group were significantly decreased(P<0.05),and the expressions of Cleaved Caspase-9 and Bax proteins were significantly increased(P<0.05).Compared with the single drug group,the combination group had a stronger regulatory effect on related proteins.[Conclusion]Xiaoyan Decoction can effectively inhibit the proliferation,induce apoptosis and S-phase block of human gastric cancer MGC-803 cells.The anti gastric cancer mechanism is related to its regulation of VEGF/PI3K/AKT signal pathway.