DKK1通过抑制MMP2、MMP9的表达影响人小细胞肺癌SBC-5细胞的增殖和侵袭能力的研究

DKK1Affects the Proliferation and Invasion of Human Small Cell Lung Cancer SBC-5 Cells by Down-regulation of MMP2 and MMP9

目的探讨下调DKK1表达对人小细胞肺癌SBC-5细胞增殖、侵袭能力的影响及机制。方法下调SBC-5细胞中DKK1表达(实验组:SBC-5-DKK1-siRNA,对照组:SBC-5-NC-siRNA),观察细胞绿色荧光比例;RT-PCR和Western-bloting验证感染效率,观察MMP2和MMP9蛋白表达的变化;平板克隆实验和细胞侵袭实验观察下调DKK1表达对SBC-5细胞增殖及侵袭能力的影响。结果细胞绿色荧光比例大于80%;DKK1表达在mRNA和蛋白水平均被下调(P<0.05),下调DKK1表达抑制了MMP2和MMP9表达;细胞克隆形成和侵袭能力均被抑制(P<0.05)。结论下调DKK1的表达,可能通过下调MMP2及MMP9的表达抑制了SBC-5细胞的增殖和侵袭能力。提示DKK1和MMP2及MMP9可能在小细胞肺癌的发生发展过程中具有重要的作用。

Objective This study aimed to examine the effects and molecular mechanisms of DKK1 on SBC-5 cells.Methods The expression of DKK1 was down-regulated by transfection with lentiviruses in SBC-5 cells(experimental group:SBC-5-DKK1-shRNA,Control group:SBC-5-NC-siRNA),the ratio of green fluorescence observed after 72 h;The expression levels of mRNA and protein identified by RT-PCR and Western-bloting;The effects of DKK1 on the proliferation and invasion were observed by colony-forming assay and Transwell methods.Results The proportion of green fluorescence was more than 80%;The expression levels of DKK1 mRNA and protein were significantly decreased(P<0.05),the expression of MMP2 and MMP9 were inhibited;The proliferation ability and invasion ability of SBC-5-DKK1-siRNA cells were inhibited(P<0.05).Conclusion Down-regulation of DKK1 can inhibit the proliferation and invasion ability of SBC-5 cells through inhibiting the expression of MMP2 and MMP9.This study indicated that DKK1,MMP2 and MMP9 may play an important role in the development of SCLC.