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

关键词:  消岩汤  MGC-803细胞  胃癌  VEGF/PI3K/AKT

Key words:  Xiaoyan Decoction  MGC-803 cell  gastric cancer  VEGF/PI3K/AKT

摘要:

[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（Westernblot）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（IC₅₀）为63.56mg/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。

Abstract:

[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 IC₅₀ 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.