雷公藤甲素通过丝氨酸-苏氨酸蛋白激酶/糖原合成酶激酶-3β通路对MGC803细胞增殖、侵袭与迁移的影响

Effect of Triptolide on Proliferation,Invasion and Migration of MGC803 Cells Through the AKT/GSK-3βPathway

目的 探讨雷公藤甲素通过丝氨酸-苏氨酸蛋白激酶/糖原合成酶激酶-3β(AKT/GSK-3β)通路对MGC803细胞增殖、侵袭与迁移的影响。方法 体外培养MGC803细胞,确定雷公藤甲素的干预剂量和干预时间。将细胞分为阴性对照组(A组,不进行处理),阳性对照组(B组,10μmol/L顺铂),雷公藤甲素高、中、低剂量组(C组、D组、E组)。干预48 h,采用Transwell法检测细胞侵袭能力,采用划痕试验法检测迁移能力,采用免疫印迹(Western blot)法检测检测相关蛋白的表达。结果 随着雷公藤甲素浓度的增加,MGC803细胞增殖率降低(P <0.05);48 h时半数抑制浓度(IC50)最小,选择干预时间为48 h,C组、D组、E组的干预剂量分别为80,40,20 nmol/L。与B组比较,C组、D组、E组侵袭细胞数、细胞迁移距离及波形蛋白(vimentin),snail,p-AKT/AKT,p-GSK-3β/GSK-3β表达水平均显著升高(P <0.05),上皮钙黏素(E-cadherin)、β-联蛋白(β-catenin)表达水平均显著降低(P <0.05),且呈剂量依赖关系。结论 雷公藤甲素可通过抑制AKT/GSK-3β信号通路而抑制MGC803细胞增殖、侵袭与迁移。

Objective To investigate the effect of triptolide on the proliferation,invasion,and migration of MGC803 cells through the serine threonine protein kinase/glycogen synthase kinase 3(AKT/GSK-3β)pathway.Methods MGC803 cells were cultured in vitro to determine the intervention dose and intervention time of triptolide.The cells were divided into the negative control group(group A,without treatment),positive control group(group B,10 mol/L cisplatin),and triptolide high-,medium-,and low-dose groups(groups C,D,and E).After 48 h of intervention,Transwell assay was used to detect cell invasion ability,scratch assay was used to detect migration ability,and Western blot was used to detect the expression of related proteins.Results With the concentration of triptolide increasing,the proliferation rate of MGC803 cells decreased(P<0.05).The IC50 was the lowest at 48 h,so 48 h was selected as the intervention time.The intervention doses for groups C,D,and E were 80,40,and 20 nmol/L,respectively.Compared with those in group B,the number of the invasive cells,the cell migration distance,and the expression levels of vimentin,snail,p-AKT/AKT,and p-GSK-3β/GSK-3βall significantly increased(P<0.05),while the expression levels of E-cadherin andβ-catenin significantly decreased in groups C,D,and E(P<0.05),which showed a dose-dependent relationship.Conclusion Triptolide can inhibit MGC803 cell proliferation,invasion,and migration by inhibiting AKT/GSK-3βsignal pathways.