伊维菌素对人胃癌细胞BGC-823、MGC-803迁移和侵袭的影响及机制研究

Effects of Ivermectin on Migration and Invasion of Human Gastric Cancer Cells BGC-823 and MGC-803 and Its Mechanism

目的:研究伊维菌素对人胃癌细胞BGC-823、MGC-803迁移和侵袭能力的影响及其作用机制。方法:0、2.5、5、10、20、40μmol/L伊维菌素分别作用于BGC-823、MGC-803细胞24 h后用MTT法检测细胞抑制率,再采用Transwell小室侵袭实验观察5μmol/L伊维菌素和含0.67‰二甲基亚砜的磷酸盐缓冲液(对照组)作用24 h对BGC-823、MGC-803细胞迁移和侵袭的影响,Western blot法分别检测5、10μmol/L伊维菌素和含0.67‰二甲基亚砜的磷酸盐缓冲液(对照组)作用于BGC-823、MGC-803细胞24 h后上皮-间质转化(EMT)标记物E-cadherin、N-cadherin、Vimentin、Snail和EMT转导通路转化生长因子β(TGF-β)/Smad中TGF-β1、TGF-βR、Smad2、Smad3蛋白的表达水平。结果:伊维菌素对BGC-823、MGC-803细胞生长均有抑制作用,其细胞抑制率与其浓度呈正相关。与对照组比较,5μmol/L伊维菌素作用后BGC-823、MGC-803细胞的迁移数和侵袭数均明显减少(P<0.01或P<0.001);5、10μmol/L伊维菌素作用后BGC-823、MGC-803细胞中E-cadherin蛋白表达明显增强(P<0.05或P<0.01或P<0.001),N-cadherin、Vimentin、Snail、TGF-βR、Smad2、Smad3蛋白表达均明显减弱(P<0.05或P<0.01或P<0.001),TGF-β1蛋白仅在10μmol/L伊维菌素作用后明显减弱(P<0.05)。结论:伊维菌素能显著抑制BGC-823、MGC-803细胞的迁移和侵袭,其可能与抑制TGF-β/Smad活性从而影响EMT过程有关。

OBJECTIVE:To study the effects of ivermectin on the migration and invasion of human gastric cancer cell lines BGC-823 and MGC-803 and its mechanism.METHODS:After treated with 0,2.5,5,10,20,40μmol/L ivermectin for 24 h,inhibitory rate of human gastric cancer cell lines BGC-823 and MGC-803 were detected by MTT assay.Effects of 5μmol/L ivermectin and phosphate buffercontaining 0.67‰dimethyl sulfoxide(control group)for 24 h on the migration and invasion of gastric cancer cells BGC-823 and MGC-803 were observed by Transwell chamber invasion assay.Western blot assay was used to detect the protein expression of TGF-β1,TGF-βR,Smad2 and Smad3 in epithelial-mesenchymal transition(EMT)markers E-cadherin,N-cadherin,Vimentin,Snail and EMT transduction pathway TGF-β/smad of BGC-823 and MGC-803 cells after treated with 5,10μmol/L ivermectin and phosphate buffercontaining 0.67‰dimethyl sulfoxide(control group)for 24 h.RESULTS:Ivermectin could inhibit the growth of BGC-823 and MGC-803,inhibitory rate of it was positively correlated with its concentration.Compared with control group,the number of migration and invasion BGC-823 and MGC-803 cells were decreased significantly after treated with 5μmol/L ivermectin(P<0.01 or P<0.001);the expression of E-cadherin protein was enhanced significantly in BGC-823 and MGC-803 cells after treated with 5 and 10μmol/L ivermectin(P<0.05 or P<0.01 or P<0.001);the protein expression of N-cadherin,Vimentin,Snail,TGF-βR,Smad2 and Smad3 were decreased significantly(P<0.05,P<0.01 or P<0.001);protein expression of TGF-β1 was decreased significantly after treated with 10μmol/L ivermectin(P<0.05).CONCLUSIONS:Ivermectin can significantly inhibit the migration and invasion of gastric cancer cells BGC-823 and MGC-803,and inhibiting the biological activity of EMT by reducing the expression of TGF-β/smad pathway is one of the mechanisms that inhibit the migration and invasion of gastric cancer cells.