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目的观察siRNA敲低S100A4的表达后对胶质瘤细胞系SNB19侵袭和迁移能力的影响。方法 S100A4干扰RNA(siRNA)敲低SNB19细胞中S100A4的表达(n=3),同时设control组(空白对照组,n=3)和siRNA-NC组(阴性对照组,n=3),采用RT-PCR和western blot方法分别检测S100A4被有效敲低,划痕实验和Transwell实验分别检测细胞的迁移和侵袭能力的变化,western blot法检测基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)和E-cadherin的表达变化,倒置相差显微镜观察细胞片状伪足变化情况。结果 siRNA技术可显著下调SNB19细胞中S100A4 mRNA和蛋白的表达水平,siRNA-NC组和siRNA-S100A4组的mRNA较control组的表达量分别是0.97±0.07和0.21±0.04(P<0.01),三个组的蛋白相对表达量分别为78.12%±2.63%、77.16%±3.00%和37.95%±2.71%(P<0.01);干扰成功后,siRNA-S100A4组与control组相比,细胞的迁移和侵袭能力分别降低了46%和55%,MMP-9和MMP-2的蛋白表达水平分别下调了62%和68%,E-cadherin的表达水平上调了154%,细胞片状伪足较对照SNB19细胞相比明显变小,差异均有统计学意义(P<0.01)。结论敲低S100A4表达可降低胶质瘤细胞系SNB19的侵袭和迁移能力,提示S100A4可能成为抗胶质瘤侵袭迁移治疗的有效靶点。
Objective To investigate the effect of siRNA knockdown of S100A4 on invasion and migration of glioma cell line SNB19. Methods The expression of S100A4 in SNB19 cells was knocked down by S100A4 siRNA (n = 3). The control group (n = 3) and siRNA-NC group (n = 3) The expression of S100A4 was knocked down by RT-PCR and western blot respectively. The migration and invasion ability of S100A4 were detected by scratch assay and Transwell assay respectively. The expressions of MMP-2, MMP-9 (MMP-9) and E-cadherin were observed by inverted phase contrast microscope. Results siRNA significantly down-regulated the expression of S100A4 mRNA and protein in SNB19 cells. The mRNA levels of siRNA-NC group and siRNA-S100A4 group were 0.97 ± 0.07 and 0.21 ± 0.04 (P <0.01) Compared with the control group, the relative expression of proteins in the siRNA-S100A4 group was 78.12% ± 2.63%, 77.16% ± 3.00% and 37.95% ± 2.71%, respectively (P <0.01) Invasiveness decreased by 46% and 55% respectively, the expression of MMP-9 and MMP-2 decreased by 62% and 68%, respectively, and the expression of E-cadherin increased by 154% Cells were significantly smaller, the difference was statistically significant (P <0.01). Conclusion Knockdown of S100A4 expression can reduce the invasion and migration of glioma cell line SNB19, suggesting that S100A4 may be an effective target for anti-glioma invasion and migration.