穗花杉双黄酮对人胃癌细胞株增殖、侵袭、迁移、粘附的影响及作用机制探讨  

Regulatory effects of Amentoflavone on the proliferation,invasion,migration and adhesion of human gastric cancer cells and the underlying mechanism

作　　者：张梅芳[1] 石建[2] 吕军[2] 薛刚[2] ZHANG Meifang;SHI Jian;LV Jun;XUE Gang(Department of Pharmacy,Rugao Hospital of Traditional Chinese Medicine,Rugao,Jiangsu 226500;Department of Internal Medicine,Rugao Hospital of Traditional Chinese Medicine,Rugao,Jiangsu 226500)

机构地区：[1]江苏省如皋市中医院药剂科,江苏如皋226500 [2]江苏省如皋市中医院内科,江苏如皋226500

出　　处：《河北中医》2024年第1期113-117,共5页Hebei Journal of Traditional Chinese Medicine

基　　金：江苏省中医药科技发展计划项目(编号:MS2021062);南通市基础研究和民生科技计划指导性项目(编号:MSZ21109)。

摘　　要：目的观察研究穗花杉双黄酮(AMF)在体外对人胃癌细胞株增殖、侵袭、迁移、粘附的影响,并探讨其作用机制。方法选择BGC-823人胃癌细胞株进行相关试验,采用四甲基偶氮唑蓝(MTT)法检测AMF对BGC-823细胞增殖的影响,Transwell侵袭试验观察AMF对BGC-823细胞侵袭的影响,细胞划痕试验观察AMF对BGC-823细胞迁移的影响,细胞粘附试验观察AMF对BGC-823细胞粘附能力的影响,并采用Western bol法检测AMF对与BGC-823细胞侵袭、迁移、粘附相关的基质金属蛋白酶2(MMP-2)、MMP-9及伴有Kazal域的富含半胱氨酸的逆转诱导蛋白(RECK)表达,以及磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)通路中PI3K、p-PI3K、Akt及p-Akt蛋白表达的影响。结果MTT试验结果显示,50、100、200、400μmol/L浓度的AMF对BGC-823细胞增殖均有抑制作用,与1‰二甲基亚砜(DMSO)组及空白对照组比较差异均有统计学意义(P<0.05),且随AMF浓度的增加抑制率逐渐升高,呈现浓度依赖性关系(P<0.05)。Transwell侵袭试验结果显示,与空白对照组相比,AMF 100、200、400μmol/L浓度组侵袭过去的细胞数目均明显低于空白对照组(P<0.05),且随AMF浓度的增加侵袭细胞数目逐渐减少(P<0.05),呈显浓度依赖关系。细胞划痕试验结果显示,与空白对照组相比,AMF 100、200、400μmol/L浓度组迁移至划痕区域内的BGC-823细胞数目均少于空白对照组(P<0.05),且随着AMF浓度增加迁移细胞数目逐渐减少(P<0.05),呈浓度依赖关系。细胞粘附试验结果显示,AMF 100、200、400μmol/L浓度组的AMF作用于BGC-823细胞20、40、60、80 min后,各浓度组在各时间点粘附于纤维粘连蛋白(FN)胶上的细胞数均低于空白对照组(P<0.05),且随着AMF浓度的增加,细胞粘附抑制率越大,随着作用时间的增加,细胞粘附抑制率亦越大,呈浓度、时间依赖关系(P<0.05)。Western bolt条带分析显示,与空白对照组比较,AMF 100、200、400μmol/L浓度组的RECK蛋白表�Objective To investigate the regulatory effects of Amentoflavone(AMF)on the proliferation,invasion,migration and adhesion of human gastric cancer cells in vitro,and its underlying mechanism.Methods The human gastric cancer cell BGC-823 was used in the following experiments.The regulatory effects of AMF on the proliferation,invasion,migration and adhesion of BGC-823 cells were detected by methyl thiazolyl tetrazolium(MTT)assay,Transwell assay,wound healing assay and cell adhesion assay,respectively.Protein expressions of those associated with cell invasion,migration and adhesion,including the matrix metalloproteinase-2(MMP-2),MMP-9 and reversion-inducing-cysteine-rich protein with kazal motifs(RECK)were detected by Western blot.Similarly,protein expressions of those involved in the PI3K/Akt signaling pathway,including the phosphatidylinositol 3-kinase(PI3K),phosphorylated PI3K(p-PI3K),protein kinase B(Akt)and phosphorylated Akt(p-Akt)were detected by Western blot as well.Results The MTT assay showed that the treatment of AMF at 50,100,200 and 400μmol/L significantly inhibited proliferation in BGC-823 cells compared with those induced with 1‰dimethyl sulfoxide(DMSO)or blank control(P<0.05).The inhibitory rate of proliferation dose-dependently increased with the increase in the concentration of AMF(P<0.05).Transwell assay showed that the number of invasive cells in BGC-823 cells induced with 100,200 and 400μmol/L was significantly less than those treated with blank control(P<0.05),which was in a dose-dependent manner(P<0.05).Wound healing assay showed that the number of migratory cells in BGC-823 cells induced with 100,200 and 400μmol/L was significantly less than those treated with blank control(P<0.05),which was in a dose-dependent manner(P<0.05).Cell adhesion assay showed that the number of adhesive cells treated with 100,200 and 400μmol/L for 20,40,60 and 80 min on the fibronectin gel was significantly less than those induced with blank control(P<0.05),which was in a dose-dependent and time-dependent manne

关 键 词：胃癌:实验 穗花杉双黄酮 中药药理 

分 类 号：R273[医药卫生—中西医结合]