Syntenin基因促进胶质瘤侵袭和迁移的分子机制

摘要

目的：探讨多配体聚糖结合蛋白（Syntenin）基因表达水平对胶质瘤细胞侵袭和迁移能力的影响以及可能的分子机制。方法采用慢病毒RNA干扰技术敲低胶质母细胞瘤细胞株U-87中Syntenin基因表达水平；qPCR检测Syntenin mRNA表达水平，筛选最佳干扰序列；Transwell小室实验检测细胞的侵袭和迁移能力变化；粘附实验检测细胞的粘附能力变化；Western-Blot（WB）检测Syntenin、AKT、p-AKT和MMP-9的表达水平。结果 qPCR结果显示，干扰组Syntenin mRNA表达水平显著低于空载组（P<0.01），空载组与对照组相比无明显差异(P>0.05)。侵袭和迁移实验结果显示，干扰组细胞的侵袭和迁移能力显著低于空载组，差异均有统计学意义（P<0.01）；空载组与对照组相比无明显差异(P>0.05)。粘附实验结果显示，在各时间点，干扰组细胞的同种粘附率明显高于空载组，差异均有统计学意义（P<0.05）；异种粘附率均明显低于空载组，差异均有统计学意义（P<0.05）。WB结果显示，干扰组Syntenin、p-AKT和MMP-9的表达水平显著低于空载组，差异均有统计学意义（P<0.05），空载组与对照组相比无明显差异(P>0.05)；AKT表达水平在各组之间均无显著差异(P>0.05)。结论在胶质瘤中，Syntenin可能通过上调AKT的磷酸化水平，经过相应的信号转导通路上调MMP-9表达水平，从而促进胶质瘤细胞的侵袭和迁移。%Objective To investigate the effect of different gene expression levels of Syntenin on invasion and mi⁃gration of glioma cells and the underlying molecular mechanisms. Methods Lentiviral RNA interference was used to knockdown the expression of syntenin in U-87 cells. Real-time quantitative PCR was used to detect mRNA expression levels of syntenin . Transwell assay and adhesion assay were used to examine the invasion, migration and adhesion, re⁃spectively. Western-blot was used to detect the protein expression levels of Syntenin, AKT, p-AKT, and MMP-9. Re⁃sults The mRNA expression level of Syntenin was greatly reduced in interference group compared with empty vector group (P<0.01). The ability of invasion and migration was much lower in interference group than in empty vector group (P<0.01). However, there were no significant differences in invasion and migration between empty vector group and con⁃trol group. The adhesion ability of glioma U-87 cells was much higher in interference group than in empty vector group (P<0.05). However, when U-87 cells were seeded on 96-wells coated with HUVEC, the adhesion ability was much lower in interference group than in empty vector group(P<0.05). The protein expression levels of Syntenin, p-AKT, and MMP-9 in interference group were markedly decreased compared with empty vector group(P<0.05). There was no signif⁃icant difference in expression of AKT protein between interference group and empty vector group (P>0.05). Conclusion Syntenin may enhance the invasion and migration ability of glioma though up-regulation of p-AKT, which in turn pro⁃motes MMP-9 expression in a corresponding signal transduction pathway.