RNA干扰下调CD98hc表达对胶质瘤细胞U251增殖、转移的影响

Effect of RNA interference down-regulation of CD98hc on proliferation,migration of U251 cells

目的观察下调CD98hc对胶质瘤U251细胞体外增殖、迁移及侵袭的影响。方法应用si RNA技术干涉U251胶质瘤细胞CD98hc的表达,筛选并获得CD98hc低表达的U251细胞株(U251KD)及阴性对照细胞(U251NC)。流式细胞技术及Western blot鉴定CD98hc的表达下调情况。应用CCK-8细胞增殖实验检测两组细胞增殖;细胞划痕实验及Transwell小室分别检测两组细胞的迁移及侵袭能力;Western blot检测MMP-2、MMP-9的表达。结果与阴性对照U251NC组相比,U251KD细胞中CD98hc水平明显下调(P<0.01),U251KD细胞的增殖和迁移、侵袭能力均显著受抑(P<0.05),MMP-2、MMP-9的表达也明显降低(P<0.01)。结论下调CD98hc的表达能够明显抑制胶质瘤U251细胞的体外增殖、迁移及侵袭,其机制可能与MMP-2、MMP-9的下调相关。CD98hc分子可作为胶质瘤治疗的一个潜在靶点。

Objective To observe the effect of down-regulation of CD98 hc on proliferation, migration and invasion of U251 cells. Methods si RNA technology was used to interference CD98 hc in U251 cells. CD98 hc low expression U251cells（U251KD） and the negative control cells（U251NC） were obtained. FCM and Western blot were used to verify the down-regulation of CD98 hc. CCK-8 assay was used to test the cell proliferation of both U251 KD and U251 NC cells.Wound scratch assay and Transwell assay were conducted to determine the migration and invasion potency of U251 KD and U251 NC cells. Western blot was used to detect the expression of MMP-2 and MMP-9. Results Compared to that of U251 NC cells, CD98 hc was down-regulated significantly in U251 KD cells（P〈0.01）. The cell proliferation, migration and invasion potency were significantly inhibited in U251 KD cells（P〈0.05）; suppressing CD98 hc expression also down-regulated the expression of MMP-2 and MMP-9（P〈0.01）. Conclusion Knocking down of CD98 hc inhibits the proliferation, migration and invasion of U251 cells in vitro, and the mechanism may be associated with the down-regulation of MMP-2 and MMP-9. CD98 hc can act as a potential target in glioma therapy.