作者单位：315000 宁波，宁波明州医院神经外科(江汉清)；510630 广州，中山大学附属第三医院(罗仑，梁朝峰)

    营养缺乏自噬因子1对人脑胶质瘤U251细胞增殖侵袭能力的影响

    江汉清罗仑梁朝峰

    【摘要】目的探讨营养缺乏自噬因子1对人脑胶质瘤U251细胞增殖侵袭能力的影响。 方法 复苏培养人脑胶质瘤U251细胞，建立转染小干扰RNA(siRNA)的NAF1组(si-NAF1组)、阴性对照组(si-NC组)、未转染siRNA对照组(control组)，采用CCK-8实验检测细胞增殖能力、平板克隆形成实验检测细胞克隆形成能力、细胞划痕实验检测各组细胞迁移能力、Transwell法观察各组细胞侵袭能力变化。结果与control组比较，NAF1-siRNA干预U251细胞后，si-NAF1组NAF1的mRNA和蛋白表达水平均明显降低(P均0.05)。结论转染siRNA后NAF1表达下降，进而下调PCNA水平，抑制人脑胶质瘤U251细胞的增殖能力；同时MMP-9蛋白表达水平下降，抑制U251细胞的迁移和侵袭能力。

    【关键词】脑胶质瘤；RNA干扰；细胞增殖；肿瘤侵袭

    DOI：10.3969/g.issn.0253-9802.2015.02.004

    通讯作者，梁朝峰，E-mail：lcfjeff@163.com

    收稿日期：(2014-10-09)

    Influence of nutrient-deprivation autophagy factor-1 on the proliferation and invasion of human glioma cell line U251JiangHanqing,LuoLun，LiangChaofeng.DepartmentofNeurosurgery，NingboMingzhouHospital,Ningbo315000,China

    Correspondingauthor,LiangChaofeng,E-mail:lcfjeff@163.com

    Abstract【】ObjectiveTo investigate the effect of nutrient-deprivation autophagy factor-1 (NAF1) upon the proliferation and invasion of human glioma cell line U251. MethodsResuscitation of human glioma cell line U251 was conducted. The cultured U251 cells were divided into the si-NAF1, si-NC and control groups. CCK-8 assay was performed to assess the proliferation of U251 cells. Plate cloning formation assay was utilized to evaluate the formation of cell cloning. Wound healing assay was conducted to investigate the migration of U251 cells. Changes in the migration of U251 cells were determined by Transwell invasion assay. ResultsThe expression of NAF1 in U251 cells transfected with NAF1-siRNA at both protein and RNA levels was significantly down-regulated compared with those in si-NC and control groups (both P0.05). ConclusionsAfter transfection with siRNA, the expression of NAF1 was decreased, thereby down-regulating the expression of PCNA and inhibiting the proliferation of U251 cells. Down-regulated expression level of MMP-9 protein suppressed the proliferation and migration of U251 cells. ......