苦参碱对神经母细胞瘤细胞增殖、迁移和侵袭的影响

Effects of matrine on the proliferation,migration,and invasion of neuroblastoma cells

目的观察苦参碱对人神经母细胞瘤细胞增殖、迁移和侵袭的影响,探讨其可能的分子机制。方法实验分为AS实验组(SK-N-AS细胞给予IC50浓度的苦参碱)、AS空白组(SK-N-AS细胞正常培养)、AS对照组(SK-N-AS细胞加入等量二甲基亚砜处理)和DZ实验组(SK-N-DZ细胞给予IC50浓度的苦参碱)、DZ空白组(SK-N-DZ细胞正常培养)、DZ对照组(SK-N-DZ细胞加入等量二甲基亚砜处理)。用划痕实验和Transwell小室检测苦参碱对各组细胞迁移和侵袭能力的影响,用蛋白质印迹法检测苦参碱对各组细胞中E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)和波形蛋白(Vimentin)蛋白表达的影响。结果各实验组不同干预后,AS空白组、AS对照组、AS实验组、DZ空白组、DZ对照组和DZ实验组的迁移百分率分别为(66.32±3.12)%、(65.27±3.44)%、(23.73±0.79)%、(46.25±4.68)%、(44.15±5.60)%和(16.77±3.52)%,侵袭细胞数分别为(870.45±19.32)、(865.32±23.39)、(492.74±16.81)、(1198.10±43.71)、(1203.03±71.91)和(891.69±42.62)个,E-cadherin蛋白表达水平分别为(100.00±11.72)%、(105.65±13.11)%、(477.20±29.71)%、(100.00±12.54)%、(97.78±12.77)%和(240.53±12.23)%,N-cadherin蛋白表达水平分别为(100.00±15.44)%、(103.90±10.76)%、(43.52±9.96)%、(100.00±10.12)%、(104.95±10.49)%和(38.39±8.70)%,Vimentin蛋白表达水平分别为(100.00±9.51)%、(97.39±11.33)%、(59.13±10.25)%、(100.00±13.20)%、(96.27±11.01)%和(47.67±9.48)%。AS实验组的上述指标与AS空白组比较,在统计学上差异有统计学意义(P<0.01、P<0.001),DZ实验组的上述指标与DZ空白组比较,在统计学上差异有统计学意义(P<0.01、P<0.001)。结论苦参碱抑制人神经母细胞瘤SK-N-AS和SK-N-DZ细胞增殖、迁移和侵袭,其机制可能与抑制上皮间质转化有关。

Objective To observe the effects of matrine on the proliferation,migration,and invasion of human neuroblastoma cells,and to investigate its potential mechanism.Methods This study was divided into AS experimental group(SK-N-AS cells treated with IC_(50)concentration of matrine),AS blank group(SK-N-AS cells cultured under normal conditions),AS control group(SK-N-AS cells treated with an equal amount of dimethyl sulfoxide),DZ experimental group(SK-N-DZ cells treated with IC_(50)concentration of matrine),DZ blank group(SK-N-DZ cells cultured under normal conditions),and DZ control group(SK-N-DZ cells treated with an equal amount of dimethyl sulfoxide).Scratch assay and Transwell chamber were used to measure the effect of matrine on the migration and invasion.The expression of E-cadherin,N-cadherin and Vimentin were tested by Western blot.Results After different intervention,the migration percentages of AS blank group,AS control group,AS experimental group,DZ blank group,DZ control group and DZ experimental group were(66.32±3.12)%,(65.27±3.44)%,(23.73±0.79)%,(46.25±4.68)%,(44.15±5.60)%and(16.77±3.52)%,respectively;the number of invasive cells were 870.45±19.32,865.32±23.39,492.74±16.81,1198.10±43.71,1203.03±71.91 and 891.69±42.62,respectively;the expression levels of E-cadherin protein were(100.00±11.72)%,(105.65±13.11)%,(477.20±29.71)%,(100.00±12.54)%,(97.78±12.77)%and(240.53±12.23)%,respectively;the expression levels of N-cadherin protein were(100.00±15.44)%,(103.90±10.76)%,(43.52±9.96)%,(100.00±10.12)%,(104.95±10.49)%and(38.39±8.70)%,respectively;Vimentin protein expression levels were(100.00±9.51)%,(97.39±11.33)%,(59.13±10.25)%,(100.00±13.20)%,(96.27±11.01)%and(47.67±9.48)%,respectively.There were statistically significant differences in the above indexes between the AS group and the AS blank group(P<0.01,P<0.001),and there were statistically significant differences between the above indexes in the DZ group and the DZ blank group(P<0.01,P<0.001).Conclusion Matrine inhibits the proliferation,migration,and invasion of neuroblastoma SK-N-AS and SK-N-DZ cells,potentially through suppressing epithelial-mesenchymal transition.