ABCE1基因沉默对人膀胱癌 T24细胞增殖和迁移能力的影响

摘要

目的：本实验通过沉默ABCE1基因在人膀胱癌T24细胞中的表达，探讨ABCE1基因对细胞的生长、细胞周期及迁移等方面的作用。方法：设计及合成ABCE1的siRNA序列，LipofectamineTM 2000转染T24细胞。采用RT-PCR和Western blot法检测基因沉默后ABCE1 mRNA和蛋白的表达情况，采用流式细胞术检测T24细胞周期。并通过CCK-8增殖实验、划痕愈合实验和细胞侵袭实验评价沉默ABCE1基因对T24细胞增殖、迁移以及侵袭能力的影响。结果：沉默ABCE1基因48 h后， T24细胞ABCE1 mRNA及蛋白表达明显降低，差异有统计学意义。 T24的细胞周期被阻滞于G0／G1期，且S期的细胞数减少，差异有统计学意义。与对照组和空白组相比，CCK-8增殖实验显示，实验组T24细胞的增殖受到明显抑制。划痕愈合实验显示，实验组迁移能力显著下降，差异有统计学意义。 Transwell小室法示，实验组T24细胞的侵袭能力显著下降，差异有统计学意义。结论：特异性干扰ABCE1基因表达可抑制人膀胱癌T24细胞的迁移能力，并抑制肿瘤细胞增殖，因此，ABCE1的siRNA序列可能成为治疗膀胱癌的有效靶点。%AIM:To investigate the effect of small interfering RNA ( siRNA)-mediated ABCE1 knockdown on the survival, cell cycle and invasion of human bladder cancer cell line T24.METHODS:The siRNA against ABCE1 was constructed and transfected into the T24 cells with LipofectamineTM 2000.The expression of ABCE1 was detected by RT-PCR and Western blot.Flow cytometry was used to detect the cell cycle.The effects of ABCE1 gene silencing on prolifera-tion, migration and invasion of T24 cells were evaluated by CCK-8 assay, wound-healing assay and Transwell invasion as-say, respectively.RESULTS: Compared with control group and blank group, the mRNA and protein levels of ABCE1 were significantly decreased in experimental group after transfected with ABCE1 siRNA.The cell cycle was arrested at G0/G1 phase and the cell number in S phase was decreased in the T24 cells.Compared with control group and blank group, the proliferation of T24 cells in experimental group was inhibited significantly, and the migration and invasion abilities of T24 cells in experimental group decreased significantly.CONCLUSION:Knockdown of ABCE1 gene may decrease migration, invasion and proliferation abilities in T24 cells.