白藜芦醇对人视网膜母细胞瘤细胞侵袭能力的影响及机制研究

目的研究白藜芦醇(resveratrol,Res)对人视网膜母细胞瘤SO-RB50细胞侵袭能力的影响,探讨Res抑制视网膜母细胞瘤细胞侵袭力的相关机制。方法分别用6.25μmol·L-1、12.50μmol·L-1、25.00μmol·L-1、50.00μmol·L-1、100.00μmol·L-1Res处理视网膜母细胞瘤SO-RB50细胞,对照组加等量5g·L-1二甲基亚砜,孵育48h后,细胞划痕实验测定细胞迁移抑制率;分别用上述浓度的Res处理视网膜母细胞瘤细胞16h,Transwell小室侵袭实验测定细胞侵袭抑制率;50.00μmol·L-1Res处理细胞48h,采用RT-PCR和Western blotting方法分别测定基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)、基质金属蛋白酶组织抑制剂-1(TIMP-1)和基质金属蛋白酶组织抑制剂-2(TIMP-2)mRNA和蛋白表达。结果 6.25μmol·L-1、12.50μmol·L-1、25.00μmol·L-1、50.00μmol·L-1、100.00μmol·L-1Res可明显抑制视网膜母细胞瘤细胞的迁移和侵袭;与对照组比较,50.00μmol·L-1Res处理细胞48h后,Res处理组MMP-2和MMP-9mRNA对照组和Res处理组分别为(1.16±0.31、0.98±0.26和0.49±0.09、0.59±0.08)]及蛋白对照组和Res处理组分别为(0.97±0.24、0.78±0.18和0.46±0.07、0.29±0.06)]的表达和活性均显著下降,TIMP-1和TIMP-2mRNA对照组和Res处理组分别为(0.35±0.06、0.37±0.07和0.79±0.11、0.93±0.25)]及蛋白表达对照组和Res处理组分别为(0.36±0.07、0.32±0.06和0.89±0.15、0.78±0.13)]均显著上升(均为P<0.05)。结论 Res通过下调MMP-2和MMP-9及上调TIMP-1和TIMP-2的表达来发挥抑制视网膜母细胞瘤细胞侵袭的作用。

Effect of resveratrol on invasive ability of human retinoblastoma cell line in vitro and its mechanism

Objective To investigate the effects of resveratrol(Res) on invasive ability of human retinoblastoma SO-RB50 cell,and further discuss its mechanisms.Methods Human retinoblastoma SO-RB50 cells were incubated with Res(6.25 μmol·L-1,12.50 μmol·L-1,25.00 μmol·L-1,50.00 μmol·L-1,100.00 μmol·L-1) for 48 hours,5 g·L-1 dimethylsulfoxide was given in the control group,then the inhibition ratio of migration was tested by cell scarification test.SO-RB50 cells were incubated with Res for 16 hours,and the inhibition ratio of invasion was tested by Transwell matrigel invasion assay.The mRNA and protein levels of matrix metalloproteinase-2(MMP-2),MMP-9,tissue inhibitor of MMP-1(TIMP-1) and TIMP-2 were detected by RT-PCR and Western blotting in human retinoblastoma cells after treated with 50.00 μmol·L-1 Res for 48 hours.Results Res with 6.25 μmol·L-1,12.50 μmol·L-1,25.00 μmol·L-1,50.00 μmol·L-1,100.00 μmol·L-1 significantly inhibited the migration and invasion of retinoblastoma cells.Compared with the control group,the expression and activity of MMP-2 and MMP-9 mRNA(1.16±0.31,0.98±0.26;0.49±0.09,0.59±0.08) and protein(0.97±0.24,0.78±0.18;0.46±0.07,0.29±0.06)were down-regulated,TIMP-1 and TIMP-2 mRNA(0.35±0.06,0.37±0.07;0.79±0.11,0.93±0.25)and protein(0.36±0.07,0.32±0.06;0.89±0.15,0.78±0.13)were up-regulated markedly after treated with 50.00 μmol·L-1 Res for 48 hours(all P<0.05).Conclusion Res inhibits human retinoblastoma cell invasion by down-regulating the expression of MMP-2,MMP-9 and up-regulating the expression of TIMP-1 and TIMP-2.