GM3通过MMP-9调节B16细胞的运动性

GM3 regulates the motility of B16 cells via MMP-9

目的通过不同的方法来研究GM3对MMP-9的调节作用。并进一步验证了GM3对细胞运动性的调节。方法分别用RT-PCR,Zymography,HPTLC,western blotting和transwell的方法来检测MMP-9的mRNA表达水平,MMP-9的活性,GM3,Akt的磷酸化以及细胞的运动性。结果通过几种不同的方法来研究是否GM3参与MMP-9的调空:a.通过转染半乳糖转移酶的顺反序列来调节GM3的表达,从而初步证明GM3可以调节MMP-9的表达;b.通过外加GM3可以清楚的观察到MMP-9的表达量升高;c.神经节苷脂抑制剂D-PDMP的加入更进一步的证明了MMP-9的表达始终与GM3的表达相关;另外,通过PI3K抑制剂的试验进一步证明了GM3通过PI3K来调节MMP-9的表达。在此基础上,更进一步的证明了神经节苷脂可以调节细胞的运动性。结论GM3可以通过PI3K/Akt信号传导途径来正调控MMP-9的表达,并且调节细胞的运动性。

Objective To investigate the regulative mechanism of GM3 to MMP-9 and its function on cell motility. Methods RT-PCR, zymography, HPTLC, Western blotting and transwell method were used to determine the expression of MMP-9, the activity of MMP-9, GM3, the activity of Akt and the motility of cells respectively. Results GM3 was found to regulate MMP-9 expression based on several criteria： 1）the expression of GM3 could be regulated by transfecting cells with GalT-6 sense or anti-sense cDNA sequence to B16 cells, which resulted in MMP-9 stimulation or suppression respectively; 2） exogenously added GM3 could significantly up-regulate the expression of MMP-9;3） the expression of MMP-9 could be suppressed by DPDMP, an inhibitor of glycosphigolipid synthesis. Furthermore, the expression of MMP-9 could be regulated by GM3 via PI3K/Akt pathway. GM3 could control the motility of B16 cells. Conclusions The expression of MMP-9 can be positively regulated by GM3 via PI3K/Akt pathway, B16 cell motility can also be regulated.