石蒜碱调控G3BP1/TGF-β/Smad信号通路抑制食管癌细胞增殖

Lycorine inhibits the proliferation of esophageal cancer cells through regulating G3BP1/TGF-β/Smad signaling pathway

目的:探讨石蒜碱对食管癌细胞增殖的影响,并分析其机制。方法:细胞计数8(cell counting kit-8,CCK-8)法分析细胞增殖抑制率,Western blot法分析GTP酶激活蛋白-Src同源结构域3-结合蛋白1[GTPase activating protein(SH3 domain)binding protein 1,G3BP1]、转化生长因子-β1(transforming growth factor-β1,TGF-β1)、p-Smad2/Smad2、p-Smad3/Smad3水平。敲低或过表达G3BP1,观察其对细胞增殖抑制率以及G3BP1、TGF-β1、p-Smad2/Smad2水平的影响。裸鼠成瘤实验验证石蒜碱对裸鼠体内瘤体的瘤重、瘤体积及G3BP1、TGF-β1、p-Smad2/Smad2水平的影响。结果:2~64μmol/L石蒜碱抑制ECA109细胞增殖,石蒜碱48 h IC50为(16.21±2.35)μmol/L。4、8、16μmol/L的石蒜碱能下调G3BP1、TGF-β1、p-Smad2/Smad2、p-Smad3/Smad3的水平。敲低G3BP1的表达能明显提高石蒜碱对ECA109细胞增殖的抑制作用,促进石蒜碱降低G3BP1、TGF-β1、p-Smad2/Smad2水平(P<0.05);上调G3BP的表达能明显下调石蒜碱对ECA109细胞增殖的抑制作用,抑制石蒜碱降低G3BP1、TGF-β1、p-Smad2/Smad2水平(P<0.05)。体内实验显示,与模型对照组比较,25、50、100 mg/kg石蒜碱能降低瘤体体积、瘤体质量,下调瘤体G3BP1、TGF-β1、p-Smad2/Smad2水平(P<0.05)。结论:石蒜碱能抑制ECA109细胞的增殖,对裸鼠体内ECA109细胞移植瘤抑瘤效果明显,其机制与调控G3BP1/TGF-β/Smad信号通路有关。

Objective:To explore the effect of lycorine on the proliferation of esophageal cancer cells and analyze its mechanism.Methods:The inhibition of cells was analyzed by cell counting kit-8(CCK-8)method,and the levels of GTPase activating protein(SH3 domain)binding protein 1(G3BP1),transforming growth factor-β1(TGF-β1),p-Smad2/Smad2 and p-Smad3/Smad3 were analyzed by Western blot.After G3BP1 knockdown or overex⁃pression,the inhibition rate of cell proliferation and the levels of G3BP1,TGF-β1,p-Smad2/Smad2 were observed.The tumorigenesis experiment in nude mice verified the effects of lycorine on the tumor weight,tumor volume and the levels of G3BP1,TGF-β1,p-Smad2/Smad2 in nude mice.Results:2~64μmol/L lycorine inhibited the prolifera⁃tion of ECA109 cells,and the IC50 of lycorine was(16.21±2.35)μmol/L for 48 hours.4,8,16μmol/L lycorine could down-regulate the level of G3BP1,TGF-β1,p-Smad2/Smad2,p-Smad3/Smad3.Knocking down the ex⁃pression of G3BP1 could obviously improve the inhibitory effect of lycorine on the proliferation of ECA109 cells,and promote lycorine to reduce the levels of G3BP1,TGF-β1 and p-Smad2/Smad2(P<0.05).Up-regulating the expression of G3BP could obviously down-regulate the inhibitory effect of lycorine on the proliferation of ECA109 cells,and inhibit lycorine to decrease the levels of G3BP1,TGF-β1 and p-Smad2/Smad2(P<0.05).In vivo ex⁃periments showed compared with the model control group,25,50 and 100 mg/kg lycorine could reduce the tumor vol⁃ume and mass,and down-regulate the levels of G3BP1,TGF-β1 and p-Smad2/Smad2(P<0.05).Conclusion:Lycorine can inhibit the proliferation of ECA109 cells,and has obvious antitumor effect on transplanted ECA109 cells of nude mice in vivo.Its mechanism is related to the regulation of G3BP1/TGF-β/Smad signaling pathway.