DKK1蛋白对SBC-3细胞增殖、迁移和侵袭能力的促进作用及其机制

Effects of DKK1 protein on proliferation,migration and invasion of SBC-3 cells and their mechanisms

目的:探讨Dickkopf1(DKK1)蛋白对人小细胞肺癌(SCLC)SBC-3细胞生物学行为的影响,并阐明其机制。方法:过表达DKK1的慢病毒和对照病毒分别感染人SCLC细胞株SBC-3,获得稳定表达DKK1蛋白的细胞系(SBC-3-DKK1组)和对照细胞系(SBC-3-NC组)。采用RT-PCR和Western blotting法检测2组SBC-3细胞中DKK1 mRNA表达水平和蛋白表达量,平板克隆实验检测2组SBC-3细胞克隆形成率,Transwell实验观察2组SBC-3细胞迁移和侵袭能力,Western blotting法检测2组SBC-3细胞中MMP-9蛋白表达量。结果:慢病毒感染SBC-3细胞72 h后细胞内绿色荧光率大于80%。与SBC-3-NC组比较,SBC-3-DKK1组SBC-3细胞中DKK1 mRNA表达水平升高(P=0.004),DKK1蛋白表达量升高,细胞克隆形成率升高(P=0.0026),迁移细胞数增多(P=0.0062),侵袭细胞数增多(P=0.0214),SBC-3细胞中MMP-9蛋白表达量升高。结论:过表达DKK1蛋白可以促进SBC-3细胞的增殖、迁移和侵袭,其作有机制可能与上凋细胞中MMP-9蛋白表达有关。

Objective:To investigate the effects of Dickkopf1(DKK1)protein on the biological behaviors of SBC-3 cells of human small cell lung cancer(SCLC),and to elucidate their mechanisms.Methods:The human SCLC cell strain SBC-3 were infected with lentiviruses over-expressing DKK1 and control viruses,respectively,and the cells stably expressing DKK1 protein(SBC-3-DKK1 group)and control cells(SBC-3-NC group)were obtained.RT-PCR and Western blotting methods were used to detect the expression levels of DKK1 mRNA and the expression amount of DKK1 protein in the SBC-3 cells in two groups;plate cloning assay was used to detect the colony formation rate of the cells in two groups;Transwell assay was used to observe the abilities of migration and invasion of the SBC-3 cells in two groups;Western blotting method was used to detect the expression amounts of MMP-9 protein in the SBC-3 cells in two groups.Results:The green fluorescence rate in the SBC-3 cells after lentivirus infection for 72 h was more than 80%.Compared with SBC-3-NC group,the expression level of DKK1 mRNA(P=0.004)and the expression amount of DKK1 protein in the SBC-3 cells in SBC-3-DKK1 group were significantly increased,the colony formation rate was significantly increased(P=0.0026),the number of migration cells was significantly increased(P=0.0062),the number of invadsion cells was significantly increased(P=0.0214),and the expression amount of MMP-9 protein was significantly increased.Conclusion:Over-expression of DKK1 protein can promote the proliferation,migration and invasion of SBC-3 cells,and their mechanisms may be related to the up-regulation of MMP-9 protein expression.