甲基硒酸调控胃癌MGC-803细胞的作用机制研究

Mechanism of methylselenic acid regulating the gastric cancer MGC-803 cells

目的探讨甲基硒酸(methylseleninic acid,MSA)对胃癌MGC-803细胞增殖、侵袭迁移和凋亡的影响及其相关机制。方法将对数生长期的胃癌MGC-803细胞随机分为对照组、不同浓度(5、10、15μmol/L)MSA组、抑制剂XAV939组和激活剂SKL2001+MSA组。采用MTT法检测MSA干预MGC-803细胞24、48和72 h后的细胞活力;Transwell小室法检测细胞侵袭、迁移能力;流式细胞术检测细胞凋亡率与周期分布;采用免疫印迹法检测β-联蛋白(β-catenin)、糖原合成酶激酶-3β(glycogen synthase kinase 3β,Gsk-3β)、轴抑制蛋白2(axis inhibition protein 2,Axin2)的表达水平。结果与对照组相比,不同浓度(5、10、15μmol/L)MSA组MGC-803细胞活力、S期细胞比例、细胞侵袭、迁移数目和β-联蛋白表达水平均降低(P<0.05);MGC-803细胞凋亡率、G0/G1期细胞比例、Gsk-3β、Axin2蛋白表达量均增高(P<0.05),且均呈浓度依赖性;与对照组相比,XAV939组MGC-803细胞活力、S期细胞比例、细胞侵袭、迁移数目和β-联蛋白表达水平均显著降低(P<0.05),MGC-803细胞凋亡率、G0/G1期细胞比例、Gsk-3β、Axin2蛋白表达水平均显著增高(P<0.05);与MSA组相比,激活剂SKL2001+MSA组MGC-803细胞活力、S期细胞比例、细胞侵袭、迁移数目和β-联蛋白表达水平均增高(P<0.05),MGC-803细胞凋亡率、G0/G1期细胞比例、Gsk-3β和Axin2蛋白表达水平均降低(P<0.05)。结论MSA可通过Wnt/β-catenin信号通路抑制胃癌细胞增殖、侵袭和迁移,并诱导其凋亡。

Objective To explore the effects of methylseleninic acid(MSA)on proliferation,invasion,migration and apoptosis of gastric cancer MGC-803 cells and the related mechanisms.Methods Gastric cancer MGC-803 cells in logarithmic growth phase were randomly divided into control group,different concentrations(5,10,15μmol/L)MSA group,inhibitor XAV939 group,activator SKL2001+MSA group.MTT colorimetric assay was used to detect the cell viability of MGC-803 cells after treated with MSA for 24 h,48 h and 72 h.Transwell Chambers method was used to detect the cell invasion and migration ability.Flow cytometry was applied to detect cell apoptosis rate and cell cycle distribution.Western blot was adopted to analyse the protein expression levels ofβ-catenin,Glycogen synthase kinase-3β(Gsk-3β),Axis inhibition protein 2(Axin2).Results Compared with the control group,different concentrations of(5,10,15μmol/L)MSA group significantly inhibited the proliferation of MGC-803 cells,decreased the proportion of S-phase cells,the number of cell invasion,migration and the protein expression levels ofβ-catenin of gastric cancer MGC-803 cells(P<0.05),but promoted the G0/G1 phase cell ratio,apoptosis rate,Gsk-3βand Axin2 protein expression(P<0.05)in a dose-dependent manner.Compared with the control group,MGC-803 cell viability,S-phase cell ratio,cell invasions number,migration number and expression levels ofβ-catenin protein in XAV939 group were significantly decreased(P<0.05),and G0/G1 phase cell ratio,apoptosis rate and expression levels of Gsk-3βand Axin2 protein were greatly increased(P<0.05).Compared with the MSA group,the SKL2001+MSA group improved cell viability,S-phase cell ratio,number of cell invasion and migration,expression ofβ-catenin protein(P<0.05),and decreased G0/G1 phase cell ratio,apoptosis rate,Gsk-3βand Axin2 protein levels(P<0.05).Conclusion MSA inhibits cell proliferation,invasion and migration of gastric cancer cells and induces cell apoptosis by Wnt/β-catenin signal pathway.