丙戊酸通过GSK3β/β-catenin信号轴抑制胃癌细胞的增殖和迁移

Valproic acid inhibits proliferation and migration of human gastric cancer cells via GSK3β/β-catenin signal axis

目的探讨丙戊酸(valproic acid,VPA)对胃癌细胞增殖、迁移的影响及可能涉及的分子机制。方法以0、2、4、6和8 mmol·L^-1浓度的VPA分别处理人胃癌细胞AGS、SGC-7901、MGC-803和BGC-82324、48、72 h后,MTT法检测细胞的活性;软琼脂克隆实验检测VPA对胃癌细胞增殖的影响;划痕实验和Transwell迁移实验检测VPA对胃癌细胞迁移能力的影响;应用STITCH、STRING和Enrichr数据库预测VPA可能影响的信号通路;免疫印迹实验检测上皮间质转化和GSK3β/β-catenin信号通路等标志物的表达情况;免疫荧光实验检测上皮间质化相关标志物的表达情况。结果与对照组相比,VPA可显著抑制AGS、SGC-7901、MGC-803和BGC-823细胞的增殖,且呈浓度和时间依赖性;可抑制AGS和SGC-7901细胞的迁移能力及EMT进程(上皮标志物E-cadherin表达上调,间质标志物Vimentin、Snail表达下调);可下调p-GSK3βser9和β-catenin的蛋白表达水平。结论VPA可有效抑制胃癌细胞的增殖和迁移,其机制可能与GSK3β/β-catenin信号轴的抑制及EMT进程的逆转有关。

Aim To evaluate the efficacy of valproic acid(VPA)on the proliferation and migration of hu-man gastric cancer cells in vitro and to explorethe mo-lecular mechanisms.Methods The cytotoxicity of VPA on human gastric cancer cells AGS,SGC-7901,MGC-803 and BGC-823 treated with various concentra-tions(0 mmol·L^-1,2 mmol·L^-1,4 mmol·L^-1,6 mmol·L^-1 and 8 mmol·L^-1)of VPA for 24,48,72 hours by MTT assay.The ability of VPA to modulate gastric cancer cells proliferation was detected by soft agar colony formation assay.The motility of VPA treat-ment on gastric cancer cells was determined by wound healing assay and transwell assay.The VPA-protein interaction network and the protein-protein interaction network were analyzed using STITCH database,STRING database and Enrichr database.The expres-sion levels of the related protein including EMT process and GSK3β/β-catenin signal axis were detected by Western blot.The expression levels of EMT markers were also detected by immunofluorescence staining.Results The cell viability significantly decreased in AGS,SGC-7901,MGC-803 and BGC-823 cells treated with VPA compared with control group,showing a con-centration-and time-dependent manner.VPA could in-hibit gastric cancer cell migration and EMT process(E-cadherin protein expression significantly increased;the expressions of Vimentin and Snail decreased).The expressions of p-GSK3βser9 andβ-catenin were down-regulated in VPA treated gastric cancer cells compared with control group.Conclusions VPA can effectively suppress the proliferation and migration of gastric canc-er cells,which may be related to modulating both GSK3β/β-catenin signal axis and epithelial-mesenchy-mal transition process.