文献类型：期刊文献

中文题名：参芪抑瘤方含药血清对胃癌细胞AGS细胞周期及Wnt/β-catenin通路的影响

英文题名：Effect of Shenqi Yiliu Formula(参芪抑瘤方)Drug-Containing Serum on the Cycle of Gastric Cancer Cell AGS and Wnt/β-Catenin Pathway

作者：魏娜[1];郭崇元[2];白敏[1];安耀荣[1];张司朝[1];武李阳[1];段永强[3]

第一作者：魏娜

机构：[1]甘肃中医药大学甘肃省中药新产品创制工程实验室,甘肃省兰州市730000;[2]甘肃省中医方药挖掘与创新转化重点实验室;[3]宁夏医科大学中医学院

第一机构：甘肃中医药大学科研实验中心（甘肃省中医药标准化技术委员会秘书处）

年份：2025

卷号：66

期号：4

起止页码：399

中文期刊名：中医杂志

外文期刊名：Journal of Traditional Chinese Medicine

收录：;北大核心:【北大核心2023】；

基金：甘肃省科技计划项目(20JR10RA314);宁夏自然科学基金(2023AAC03170)。

语种：中文

中文关键词：胃癌前病变;参芪抑瘤方;胃癌细胞;细胞迁移;细胞周期;无翅型MMTV整合位点家族成员;β-连环蛋白

外文关键词：precancerous lesions of gastric cancer;Shenqi Yiliu Formula(参芪抑瘤方);gastric cancer cells;cell migration;cell cycle;Wnt family;β-catenin

摘要：目的从无翅型MMTV整合位点家族成员(Wnt)/β-连环蛋白(β-catenin)通路探讨参芪抑瘤方治疗胃癌前病变(PLGC)的可能作用机制。方法采用CCK-8法检测胃癌细胞AGS细胞存活率筛选参芪抑瘤方含药血清干预时间及Wnt/β-catenin通路抑制剂XAV939干预浓度。实验将AGS细胞分为胃癌细胞组(15%空白血清)、抑制剂组(筛选浓度的XAV939)、参芪抑瘤方高剂量组(12%参芪抑瘤方含药血清+3%空白血清)、参芪抑瘤方中剂量组(6%参芪抑瘤方含药血清+9%空白血清)、参芪抑瘤方低剂量组(3%参芪抑瘤方含药血清+12%空白血清),每组设3个复孔。分别培养24 h、48 h通过细胞划痕实验检测细胞迁移、侵袭能力;培养筛选干预时间后流式细胞术检测细胞周期,荧光免疫法检测增殖细胞核抗原(Ki67)蛋白水平,蛋白免疫印记法检测Wnt、β-catenin、糖原合成酶激酶3β(GSK-3β)、核内T细胞特异性因子(TCF)蛋白水平,实时荧光定量PCR法检测Wnt、β-catenin、GSK-3β、TCF mRNA表达。结果筛选出参芪抑瘤方含药血清干预时间为48 h和XAV939干预浓度为20μmol/L用于后续实验。与胃癌细胞组同时间点比较,除24 h参芪抑瘤方低剂量组之外,参芪抑瘤方各剂量组24 h、48 h细胞划痕迁移率均降低(P<0.05)。与参芪抑瘤方低剂量组同时间点比较,参芪抑瘤方高、中剂量组24 h及参芪抑瘤方高剂量组48 h细胞划痕迁移率均降低(P<0.05)。与胃癌细胞组比较,参芪抑瘤方高剂量组和抑制剂组Wnt、β-catenin、TCF蛋白水平及其相应的mRNA表达降低,Ki67蛋白水平及细胞周期S期、G2期比例亦降低,GSK-3β蛋白水平、GSK-3βmRNA表达及G1期比例升高(P<0.05)。与抑制剂组比较,参芪抑瘤方高剂量组G1期比例降低、G2期比例升高(P<0.05),而Wnt、β-catenin蛋白水平其相应的mRNA表达差异无统计学意义(P>0.05)。结论参芪抑瘤方含药血清可以抑制胃癌细胞AGS的迁移、侵袭能力,将细胞周期阻滞在G1期,其机制可能与调控Wnt/β-catenin通路有关。
Objective To investigate the potential mechanism of Shenqi Yiliu Formula(参芪抑瘤方)in treating precancerous lesions of gastric cancer(PLGC)by the Wnt/β-catenin signaling pathway.Methods The CCK-8assay was used to determine the optimal intervention time for Shenqi Yiliu Formula drug-containing serum and the concentration of the Wnt/β-catenin pathway inhibitor XAV939 depends on the survival rate of AGS gastric cancer cell line.AGS cells were divided into the gastric cancer cell group(15%blank serum),inhibitor group(selected concentration of XAV939),high-dose Shenqi Yiliu Formula group(12%Shenqi Yiliu Formula drug-containing serum+3%blank serum),medium-dose Shenqi Yiliu Formula group(6%Shenqi Yiliu Formula drug-containing serum+9%blank serum),and low-dose Shenqi Yiliu Formula group(3%Shenqi Yiliu Formula drug-containing serum+12%blank serum).Each group was tested in triplicate.After culturing for 24 and 48 hours,cell migration and invasion were assessed by scratch assays;after a selected intervention period(48 hours),cell cycle distribution was analyzed using flow cytometry,Ki67 protein levels were detected by immunofluorescence,the protein levels of Wnt,β-catenin,GSK-3β,and intranuclear T-cell specific factor(TCF)were measured by the protein immunoblotting assay,and the mRNA expressions of these above factors were determined by quantitative real-time PCR.Results The optimal intervention time for Shenqi Yiliu Formula drug-containing serum was determined to be 48 hours,and the effective concentration of XAV939 was 20μmol/L.Compared with the gastric cancer cell group,Shenqi Yiliu Formula at all doses reduced the cell migration rate at 24 and 48 hours(P<0.05),except for the low-dose group at 24 hours.Compared to the low-dose group at corresponding time points,high-and medium-dose Shenqi Yiliu Formula groups showed significantly reduced migration rates,particularly the high-dose group at 48 hours(P<0.05).Compared with the gastric cancer cell group,the high-dose Shenqi Yiliu Formula and inhibitor groups exhibited reduced protein and mRNA levels of Wnt,β-catenin,and TCF,along with reduced Ki67 protein levels and a decreased proportion of cells in the S and G2 phases of the cell cycle,but GSK-3βprotein levels,GSK-3βmRNA expression,and the proportion of cells in the G1 phase increased(P<0.05).Compared to the inhibitor group,the high-dose Shenqi Yiliu Formula group showed a decreased proportion of G1-phase cells and an increased proportion of G2-phase cells(P<0.05),although differences in Wnt andβ-catenin protein levels and mRNA expressions were not statistically significant(P>0.05).Conclusion Shenqi Yiliu Formula drug-containing serum inhibits the migration and invasion of gastric cancer AGS cells and block the cell cycle at G1 phase,and its underlying mechanism may be related to the regulation of the Wnt/β-catenin signaling pathway.