毛蕊花苷对胃癌细胞生物学行为及上皮间质转化的影响及机制  

Effects and mechanisms of verproside on biological behavior and epithelial-mesenchymal transition of gastric cancer cells

作　　者：张萌 李小环[2] 祁海莉 贵永贤[3] 李彩芳[1] ZHANG Meng;LI Xiaohuan;QI Haili;GUI Yongxian;LI Caifang(Department of Clinical Pharmacology,Xinriang Central Hospital,Xinriang,Henan 453000,China;Department of Endoscopic Treatment,Xinriang Central Hospital,Xinziang,Henan 453000,China;Department of Medical Oncology,Xinxiang Central Hospital,Xinriang,Henan 453000,China)

机构地区：[1]新乡市中心医院临床药学科,河南新乡乡453000 [2]新乡市中心医院内镜诊疗部,河南新乡453000 [3]新乡市中心医院肿瘤内科,河南新乡453000

出　　处：《中华实用诊断与治疗杂志》2024年第11期1086-1092,共7页Journal of Chinese Practical Diagnosis and Therapy

基　　金：河南省医学科技攻关计划项目(LHGJ20230882)。

摘　　要：目的观察毛蕊花苷(VER)对胃癌细胞增殖、迁移、侵袭及上皮-间质转化的影响,探讨其与PTEN/Akt信号通路的关系。方法取对数生长期胃癌SGC-7901、MGC80-3、MKN45、AGS细胞,应用浓度为0.5、1、10、25、50、100μmol/L的VER处理48 h,采用CCK-8法检测细胞增殖的吸光度值,计算细胞增殖抑制率。选择细胞增殖抑制率最高的MGC80-3细胞,以及25、50、100μmol/L浓度的VER进行后续试验。取对数生长期MGC80-3细胞分为对照组(正常培养)、VER低浓度组(25μmol/L浓度的VER处理)、VER中浓度组(50μmol/L浓度的VER处理)、VER高浓度组(100μmol/L浓度的VER处理)、VER高浓度+SF1670组(100μmol/L浓度的VER+10μmol/L的PTEN抑制剂SF1670处理)。处理48 h取5组细胞,采用EdU荧光探针法检测细胞增殖,采用Transwell小室实验检测细胞迁移和侵袭,采用Western blot法检测PTEN、p-Akt/Akt、增殖细胞核抗原(PCNA)、基质金属蛋白酶-9(MMP-9)、N-cadherin、E-cadherin蛋白相对表达量。结果5组细胞EdU阳性率、迁移细胞数、侵袭细胞数及PTEN、p-Akt/Akt、PCNA、MMP-9、N-cadherin、E-cadherin蛋白相对表达量比较差异均有统计学意义(F=28.657~68.213,P均<0.001)。VER低、中、高浓度组EdU阳性率[(28.75±3.08)%、(21.64±2.37)%、(15.37±1.59)%]、p-Akt/Akt(0.79±0.08、0.53±0.06、0.35±0.04)及PCNA(0.86±0.09、0.67±0.07、0.42±0.05)、MMP-9(0.87±0.09、0.71±0.08、0.53±0.05)、N-cadherin(0.72±0.08、0.54±0.06、0.35±0.04)蛋白相对表达量均低于对照组[(45.39±5.61)%、0.96±0.10、1.15±0.13、1.02±0.11、0.92±0.09](P<0.05),迁移细胞数[(175.39±19.47)、(137.29±15.91)、(103.96±11.57)个]、侵袭细胞数[(119.47±13.82)、(93.68±10.37)、(71.09±8.35)个]均少于对照组[(216.53±23.36)、(153.72±16.48)个](P<0.05),PTEN(0.51±0.07、0.69±0.09、0.91±0.10)、E-cadherin(0.65±0.08、0.80±0.08、1.01±0.11)蛋白相对表达量均高于对照组(0.38±0.04、0.43±0.04)(P<0.05),其中3组EdU阳性率、迁移�Objective To observe the effects of verproside(VER)on proliferation,migration,invasion and epithelial-mesenchymal transition of gastric cancer cells,and to investigate its relationship with PTEN/Akt signaling pathway.Methods The log-phase gastric cancer cells SGC-7901,MGC80-3,MKN45 and AGS were treated with VER of 0,5,1,10,25,50 and 100μmol/L for 48 h.The optical density of cell proliferation was measured by CCK-8 assay,and the inhibition rate of cell proliferation was calculated.The MGC80-3 cells with the highest inhibition rate of cell proliferation and 25,50 and 100μmol/L VER were selected for subsequent experiments.The log-phase MGC80-3 cells were divided into control group(normal culture),low-dose VER group(treated with 25μmol/L VER),medium-dose VER group(treated with 50μmol/L VER),high-dose VER group(treated with 100μmol/L VER),and high-dose VER+SF1670 group(treated with 100μmol/L VER+10μmol/L PTEN inhibitor SF1670).After 48 h of treatment,the cells from all five groups were harvested.The cell proliferation was detected by EdU fluorescence probe method,the cell migration and invasion were detected by Trans well chamber assay,and the relative expressions of PTEN,p-Akt/Akt,proliferating cell nuclear antigen(PCNA),matrix metalloproteinase-9(MMP-9),N-cadherin and E-cadherin proteins were detected by Western blot.Results There were significant differences in the EdU positive rate,number of migration cells,number of invasion cells and relative expressions of PTEN,p-Akt/Akt,PCNA,MMP-9,N-cadherin and E-cadherin proteins among five groups(F=28.657-68.213,all P values<0.001).The EdU positive rates[(28.75±3.08)%,(21,64±2.37)%,(15.37±1.59)%],and relative expressions of p-Akt/Akt(0.79±0.08,0.53±0.06,0.35±0.04),PCNA(0.86±0.09,0.67±0.07,0.42±0.05),MMP-9(0.87±0.09,0.71±0.08,0.53±0.05),and N-cadherin(0.72±0.08,0.54±0.06,0.35±0.04)in low-,medium-and high-dose VER groups were lower than those in trontrol group[(45,39±5,61)%,0.96±0.10,1.15±0.13,1,02±0.11,0.92±0.09](P<0.05),the numbers of migration ce

关 键 词：胃癌 毛蕊花苷 上皮-间质转化 PTEN/Akt信号通路 MGC80-3细胞 

分 类 号：R96[医药卫生—药理学]