siRNA沉默CD99基因对人髓母细胞瘤Daoy细胞株迁移和侵袭能力的影响

Effect of siRNA Silencing CD99 Gene on Migration and Invasion of Human Medulloblastoma Daoy Cell Line

目的研究细胞黏附分子CD99对人髓母细胞瘤Daoy细胞株迁移及侵袭能力的影响及可能机制。方法采用化学合成CD99基因的小干扰RNA(CD99 siRNA)下调该基因表达,采用实时荧光定量聚合酶链式反应(quantitative real-time polymerase chain reaction, qPCR)检测转染后的细胞内CD99、基质金属蛋白酶2(matirx metallo-proteinases 2,MMP 2)和MMP 9的表达,通过细胞划痕实验和Transwell侵袭实验分别检测CD99对Daoy细胞迁移和侵袭的影响。结果 CD99-siRNA-2#能有效降低Daoy细胞株中CD99 mRNA和蛋白水平的表达,沉默后细胞的迁移和侵袭能力均显著降低,进一步检测发现MMP 9的mRNA水平亦明显下调,与对照组比较差异有统计学意义(P<0.01)。结论 CD99基因对髓母细胞瘤Daoy细胞的迁移和侵袭能力相关,CD99可能通过影响MMP 9的表达参与其中,有望成为髓母细胞瘤治疗的靶点。

Objective To investigate the effects of cell adhesion molecule CD99 gene on the migration and invasion of human medulloblastoma Daoy cell line and the possible mechanism.Methods The expression of CD99 gene was down-regulated by siRNA.The expressions of CD99 after transfected,matrix metalloproteinase 2(MMP 2)and MMP 9 were evaluated by quantitative real-time polymerase chain reaction(qPCR).The effect of CD99 silencing on the migration and invasion of Daoy cell line was evaluate by wound healing assay and Transwell assay,respectively.Results Compared with the control group,the expressions of CD99 mRNA and protein were significantly down-regulated in Daoy cell line transfected of CD99-targeting siRNA-2#(P<0.01),as well as the expressions of MMP 9 mRNA.The migration and invasion of Daoy cells transfected of CD99-targeting siRNA-2#were lower than the control group(P<0.01).Conclusion CD99 gene may decrease migration and invasion of Daoy cells through influencing the expressions of MMP 9.It is expected to be a target for the treatment of medulloblastoma.