摘要
目的:观察不同浓度胃复方水提物(WFF)对人胃癌HGC-27细胞的增殖抑制作用,进一步探讨WFF调控HIF-1α/VEGF信号通路的作用机制。方法:采用不同浓度(1.2 mg/mL、1.3 mg/mL、1.4 mg/mL、1.5 mg/mL、1.6 mg/mL、1.7 mg/mL、1.8 mg/mL)的WFF,CCK-8法寻找最佳干预浓度并观察不同的干预组别[非特异性对照组(NC组)、WFF组、WFF+HIF-1α过表达质粒组、WFF+2-MeOE2 (HIF-1α抑制剂)组、HIF-1α过表达质粒组、2-MeOE2组、甲磺酸阿帕替尼组]对HGC-27细胞增殖的影响;运用血管生成实验观察不同组别对胃癌细胞血管生成能力的影响;进一步采用Western blot法观察不同组别[NC组、WFF组、WFF+HIF-1α过表达质粒组、WFF+2-MeOE2组(HIF-1α抑制剂)、HIF-1α过表达质粒组、2-MeOE2组]对HIF-1α/VEGF信号通路相关蛋白表达的影响。结果:CCK-8法验证了WFF与NC组比较,对HGC-27细胞增殖具有抑制作用,差异有统计学意义(P<0.01),且呈剂量依赖性。与NC组比较,WFF组、WFF+HIF-1α过表达质粒组、WFF+2-MeOE2 (HIF-1α抑制剂)组、2-MeOE2组、甲磺酸阿帕替尼组对HGC-27细胞增殖具有抑制作用,差异有统计学意义(P<0.01);与WFF组比较,WFF+HIF-1α过表达质粒组细胞活性增高,差异有统计学意义(P<0.01),WFF+2-MeOE2组活性降低,差异有统计学意义(P<0.01)。与HIF-1α过表达质粒组比较,WFF+HIF-1α过表达质粒组有效抑制了HGC-27细胞增殖,差异有统计学意义(P<0.01);与2-MeOE2组比较,WFF+2-MeOE2组有效抑制了HGC-27细胞增殖,差异有统计学意义(P<0.01)。血管生成实验验证了WFF有效降低了人胃癌HGC-27细胞血管生成能力,除HIF-1α过表达质粒组外,其余各组与NC组比较,均降低了HGC-27细胞血管生成能力,差异有统计学意义(P<0.05,P<0.01)。与WFF组比较,WFF+HIF-1α过表达质粒组,成管数目、交叉点、网眼数及长度增多,差异有统计学意义(P<0.01),WFF+2-MeOE2组,成管数目、交叉点、网眼数及长度减少,差异有统计学意义(P<0.0
Objective:To observe the inhibitory effect of different concentrations of Weifufang water extract(WFF)on the proliferation of human gastric cancer HGC-27 cells,and to further explore the mechanism of the regulation of HIF-1α/VEGF signaling pathway by WFF.Methods:Used different concentrations(1.2 mg/mL,1.3 mg/mL,1.4 mg/mL,1.5 mg/mL,1.6 mg/mL,1.7 mg/mL,1.8 mg/mL)of WFF.CCK-8 method was used to find the optimal intervention concentration and observe the effects of different intervention groups[non-specific control group(NC group),WFF group,WFF+HIF-1αoverexpression plasmid group,WFF+2-MeOE2(HIF-1αinhibitor)group,HIF-1αoverexpression plasmid group,2-MeOE2 group,Apatinib Mesylate group]on proliferation of HGC-27 cells.Angiogenesis experiment was used to observe the effect of different groups on the angiogenesis ability of gastric cancer cells.Western blot method was used to observe the effects of different groups[NC group,WFF group,WFF+HIF-1αoverexpression plasmid group,WFF+2-MeOE2(HIF-1αinhibitor),HIF-1αoverexpression plasmid group,2-MeOE2 group]on the expression of HIF-1α/VEGF signaling pathway-related proteins.Results:CCK-8 method verified that compared with NC group,WFF inhibited HGC-27 cell proliferation,the difference was significant(P<0.01),and it was dose-dependent.Compared with NC group,WFF group,WFF+HIF-1αoverexpression plasmid group,WFF+2-MeOE2(HIF-1αinhibitor)group,2-MeOE2 group,Apatinib Mesylate group had a statistically significant inhibitory effect on the proliferation of HGC-27 cells,the difference was significant(P<0.01).Compared with WFF group alone,the activity of cells in WFF+HIF-1αoverexpression plasmid group increased,the difference was statistically significant(P<0.01),and the activity in WFF+2-MeOE2 group decreased,the difference was statistically significant(P<0.01).Compared with HIF-1αoverexpression plasmid group,WFF+HIF-1αoverexpression plasmid group effectively inhibited the proliferation of HGC-27 cells,the difference was statistically significant(P<0.01).Compared with 2-MeOE2
作者
孙蕊
李东芳
SUN Rui;LI Dongfang(Hu'nan Cancer Hospital,Changsha Hu'nan 410006,China)
出处
《新中医》
CAS
2023年第17期157-164,共8页
New Chinese Medicine
基金
湖南省中医药管理局科研计划项目重点课题(C2022007)
湖南省“十四五”第一批中医药领军人才和学科带头人培养项目(湘中医药[2022]4号)
湖南省2022年科药联合基金项目(2022JJ80077)。
