摘要目的 观察核因子IA (NFIA)在胶质瘤中的表达及对胶质瘤生物学功能的影响.方法 采用免疫组织化学法检测49例胶质瘤组织和10例正常脑组织中NFIA的表达.使用RNA干扰技术处理胶质瘤U251细胞,采用实时荧光定量PCR(Real-time PCR)和Western blot检测NFIA表达水平的变化,分别采用噻唑蓝法(MTT)、划痕实验及Transwell侵袭实验,检测敲低NFIA表达对胶质瘤细胞增殖、侵袭和迁移能力的变化;Western blot检测相关蛋白表达变化.结果 免疫组织化学检测结果显示NFIA在胶质瘤组织中的表达明显高于正常脑组织.下调胶质瘤细胞中NFIA表达后:MTT实验显示阴性对照组(1.367 ±0.021)与空白对照组(1.330 ±0.046)细胞生长差异无统计学意义(P=0.500),NFIA干扰组(0.823±0.096)细胞生长明显减慢,与阴性对照组与空白对照组比较,差异均有统计学意义(P=0.001).划痕实验显示NFIA干扰组细胞迁移距离(57.9±6.9) μm明显低于阴性对照组[(125.4±10.7) μm]和空白对照组[(111.5±5.9)μm],差异有统计学意义(P=0.001),Transwell实验显示RNA干扰组穿膜细胞数[(33.3±3.2)个]低于阴性对照组和空白对照组[(80.3±4.5)、(87.7±5.0)个],差异有统计学意义(P =0.001).Western blot结果显示,NFIA干扰组Ezrin蛋白相对表达(0.33 ±0.05)显著低于阴性对照组(1.08±0.08)与空白对照组(1.00±0.05),p21蛋白相对表达(1.02±0.08)显著低于阴性对照组(0.25±0.04)与空白对照组(0.29±0.03),差异均有统计学意义(P=0.001).结论 NFIA在胶质瘤组织中高表达,下调NFIA表达可抑制胶质瘤细胞增殖、侵袭和迁移能力.

abstractsObjective To investigate the effects of nuclear factor IA (NFIA) on proliferation,invasiveness and migration of the glioma.Methods The expression of NFIA in 49 cases of glioma tissues and 10 cases of normal brain tissues was detected by immumohistochemical staining.lines.Small interfering RNA (siRNA) targeting NFIA was synthesized and transfected into glioma cell line U251 cells,and the expression of NFIA were detected in the transfected glioma cells by Real-time PCR and Western blotting assays.Cell viability was measured by methyl thiazol tetrazolium (MTT) assay,Transwell assay and wound healing assay were used to determine the invasion and migration of the cells,respectively.Results Immunohistochemical staining revealed that the expression was significantly up-regulated in glioma tissues as compared with normal brain tissues.The mRNA and protein expression levels of NFIA in RNAi group (siRNA2 and siRNA3) (0.44 ± 0.03,0.26 ± 0.03) were significantly lower than those in the negative control group (1.06 ±0.06) and blank control group (1.08 ± 0.03) (P =0.001).MTT assay showed that the ability of cell proliferation at 48 hours in RNAi group was significantly declined than those in negative control group and blank control group (P =0.001),while there was no significant difference in the growth rate between negative control group (1.367 ± 0.021) and blank control group (1.330 ± 0.046)(P =0.500);Wound healing assay indicated that the scrape wound recovered significantly lower in the NFIA RNAi group (57.9 ±6.9) μm than that in the negative control group [(125.4 ± 10.7) μm] and blank control group [(111.5 ± 5.9) μm,P =0.001];Transwell assay showed that the invasiveness capability of cells in NFIA RNAi group [(33.3 ± 3.2) cells] was significantly decreased as compared with thatin the negative control group [(80.3 ± 4.5) cells] and blank control group [(87.7 ± 5.0) cells,P =0.001].The expression of Ezrin in NFIA RNAi group (0.33 ±0.05) was significantly decreased as compared with that in the negative control group (1.08 ± 0.08) and blank control group (1.00 ± 0.05) (P =0.001).p21 expression level in NFIA RNAi group (1.02 ± 0.08) was significantly increased as compared with that in the negative control group (0.25 ± 0.04) and blank control group (0.29 ± 0.03) (P =0.001).Conclusion NFIA in the glioma tissues are significantly increased as compared with those in the normal brain tissues.Down-regulation of NFIA inhibited glioma cell proliferation,invasion and migration ability.