ATP1A1基因沉默抑制人胶质瘤细胞U251侵袭

目的探讨沉默Na+/K+ATP酶A1亚基(ATP1A1)对人U251胶质瘤细胞系侵袭能力的影响及其机制。方法用shRNA-ATP1A1慢病毒感染人U251胶质瘤细胞,RT-q PCR及Western blot分别检测ATP1A1 mRNA和蛋白的表达;MTT法检测细胞体外的增殖;细胞划痕实验及Transwell小室检测细胞的迁移及侵袭能力;Western blot检测基质金属蛋白酶2/9(MMP-2/9)的表达。结果沉默ATP1A1细胞的ATP1A1 mRNA和蛋白表达均受到了明显抑制;细胞的增殖和迁移、侵袭能力也显著受抑(P0.05);MMP-2和MMP-9的表达也明显降低(P0.05)。结论靶向ATP1A1干扰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,ATP1A1可能作为胶质瘤治疗的一个潜在靶点。

ATP1A1 gene silencing inhibits the invasion of human glioma cells U251 in vitro

Objective To investigate the effect and mechanism of silencing ATP1A1 gene on invasion ability of human U251 glioma cells.Methods The human U251 glioma cells were infected with lentivirus expressing shRNA-ATP1A1.The mRNA and protein expression of ATP1A1 in U251 glioma cells was detected by RT-qPCR and Western blot,respectively.The proliferation of U251 glioma cells was determined by MTT assay.The migration and invasion ability were detected by cell scratch test and Transwell chamber.The protein expression of matrix metallo proteinase 2 (MMP-2) and MMP-9 in U251 glioma cells were detected by Western blot.Results The mRNA and protein expression of ATP1A1 in the silence group were significantly inhibited,The ability of proliferation, migration and invasion were also significantly inhibited (P<0.05),The expression of MMP-2 and MMP-9 were also significantly reduced,there was a significant difference(P<0.05).Conclusions RNA interference targeting ATP1A1 gene can significantly inhibit the proliferation, migration and invasion of glioma U251 cells.The mechanism might be related to the down-reguLation of MMP-9 and MMP-2.ATP1A1 can be used as a potential target for the treatment of glioma.