海藻糖A抑制骨肉瘤细胞增殖信号通路的研究

Inhibitory effects of glaucocalyxin A on proliferation signaling pathway of osteosarcoma cells

目的探讨海藻糖A对骨肉瘤细胞增殖转移能力抑制作用及其机制。方法将骨肉瘤细胞(上海传秋生物科技有限公司提供)分别分为4组。空白组细胞不使用药物干预,低、中、高剂量组细胞分别采用含0.25、0.50、1.00μmol/L海藻糖A的培养基培养干预不同时间,检测细胞增殖、转移以及相关蛋白表达。然后分别在4组细胞中5.00μmol/L转化生子因子-β(TGF-β)培养24 h再次检测细胞存活率、转移以及相关蛋白表达。采用方差检验及t检验。结果空白组细胞增殖抑制率(0%)、转移抑制率(0%)显著低于药物干预组[(62.45±23.54)%、(84.15±20.10)%,t=14.111、7.959,P<0.05],差异有统计学意义,N-钙黏蛋白(N-cadherin)、编码锌指蛋白转录因子(Snail)、锌指转录因子(Slug)、波形蛋白(Vimentin)(100%、100%、100%、100%)表达量显著高于治疗组[低剂量组:(72.56±12.25)%、(65.34±10.56)%、(70.41±16.96)%、(71.45±18.96)%;中剂量组:(62.36±10.58)%、(53.47±9.85)%、(51.89±18.47)%、(42.47±9.56)%;高剂量组:(21.17±8.56)%、(19.19±5.56)%、(17.49±6.25)%、(10.77±3.25)%,t=8.246、9.163、8.035、7.441,P<0.05],差异均有统计学意义;各干预组细胞随着药物浓度升高存活率(15.25%、28.22%、39.45%)、转移抑制率显著升高(10.36%、18.45%、45.25%,t=8.644、8.603,P<0.05),差异有统计学意义,N-cadherin、Snail、Slug、Vimentin表达量显著下降。各组细胞在接受TGF-β干预后增殖抑制率和转移抑制率均显著下降,上述蛋白表达量显著升高。结论海藻糖A可以有效抑制骨肉瘤细胞增殖转移,其作用与抑制TGF-β/Smad2/3信号通路相关。

Objective To investigate the inhibitory effect of glaucocalyxin A on proliferation and metastasis of osteosarcoma cells and its mechanism.Methods Osteosarcoma cells were divided into four groups.No drug intervention was used in the blank group.Cells in low,medium and high dose groups were treated with 0.25,0.50 and 1.00μmol/L glaucocalyxin A at different time points,respectively.Cell proliferation,metastasis and related protein expression were detected.Then,5.00μmol/L transforming growth factor-β(TGF-β)was cultured in four groups of cells for 24 h to detect cell survival,metastasis and related protein expression.Results The cell proliferation inhibition rate(0%)and metastasis inhibition rate(0%)in blank group were significantly lower than those in drug intervention groups[(62.45±23.54)%,(84.15±20.10)%,t=14.111,7.959,P<0.05].The expression levels of N-cadherin,Snail,Slug and Vimentin(100%,100%,100%,100%)in blank group were significantly higher than those in treatment groups[Low dose group:(72.56±12.25)%、(65.34±10.56)%、(70.41±16.96)%、(71.45±18.96)%;Medium dose group:(62.36±10.58)%、(53.47±9.85)%、(51.89±18.47)%、(42.47±9.56)%;High dose group:(21.17±8.56)%、(19.19±5.56)%、(17.49±6.25)%、(10.77±3.25)%,t=8.246、9.163、8.035、7.441,P<0.05].The cell survival rate(15.25%,28.22%,39.45%)and metastasis inhibition in intervention groups increased with increases in drug concentrations.The metastasis inhibition rates(10.36%,18.45%,45.25%)increased significantly(t=8.644,8.603,P<0.05),and the expression levels of N-cadherin,Snail,Slug and Vimentin decreased significantly.The inhibitory rate of proliferation and metastasis of cells in each group decreased significantly after receiving the intervention of TGF-β,and the expression of the above-mentioned proteins increased significantly.Conclusion Glaucocalyxin-A can effectively inhibit the proliferation and metastasis of osteosarcoma cells,and its effect is related to the inhibition of TGF-β/Smad2/3 signaling pathway.