胶质瘤中PIN1-Nanog相关通路表达的研究  

Relation of peptidyl-prolyi isomerase 1 and Nanog expressions in human gliomas

作　　者：倪升远 牛朝诗 杨洋 张翔宇 

机构地区：[1]安徽医科大学附属省立医院神经外科、脑功能与脑疾病安徽省重点实验室、安徽省脑立体定向神经外科研究所,合肥230001

出　　处：《中华神经医学杂志》2015年第3期227-232,共6页Chinese Journal of Neuromedicine

基　　金：国家自然科学基金（81172407）

摘　　要：目的 观察肽基脯氨酰异构酶1（PIN1）、Nanog在胶质瘤组织中的表达以及PIN1抑制剂PiB对U87细胞增殖能力及PIN1、Nanog表达的影响. 方法 选取安徽医科大学附属省立医院神经外科自2013年3月至2014年8月间手术切除的84例胶质瘤和10例正常脑组织标本,免疫组化染色检测标本PIN1、Nanog蛋白的表达.体外常规培养U87胶质瘤细胞,用0.2、0.5、1.0、2.0 μg/mL PiB分别作用24、48 h,MTT检测PiB对细胞增殖的抑制作用并计算抑制率;以1.0μmol/L PiB处理U87细胞24 h为1.0 μmol/L PiB组,未经任何处理的U87细胞作为对照组,免疫荧光双染检测细胞PIN1、Nanog的表达,RT-PCR和Westem blotting分别检测细胞PIN1、Nanog mRNA和蛋白的表达. 结果 WHOⅡ级、Ⅲ级、Ⅳ级胶质瘤中PIN1、Nanog高表达率不同,差异有统计学意义（P＜0.05）,且胶质瘤级别越高,PIN1、Nanog高表达率越高.MTT检测显示PiB对U87细胞的增殖有抑制作用,且呈时间和剂量依赖性.与对照组比较,1.0 μmol/L PiB组PIN1、Nanog及PIN1/Nanog阳性细胞百分比降低,差异有统计学意义（P＜0.05）;对照组和1.0 μmol/L PiB组细胞PIN1 mRNA的表达量分别为1.82±0.03和0.94±0.20,Nanog mRNA的表达量分别为1.47±0.04和0.82±0.19,差异均有统计学意义（P＜0.05）;对照组和1.0 μmol/L PiB组细胞PIN1蛋白表达量分别为2.96±0.05和1.15±0.26,Nanog蛋白表达量分别为1.52±0.16和0.75±0.05,差异均有统计学意义（P＜0.05）. 结论 PIN1、Nanog在胶质瘤的发生发展中起重要作用,且与肿瘤的恶性程度相关.PIN1可参与调控Nanog,两者之间存在相关通路,下调PIN1-Nanog通路后,胶质瘤细胞增殖能力下降.Objective To obsevre the peptidyl-prolyi isomerase 1 （PIN1） and Nanog expreesions in glioma tissues and the effect of PIN1 inhibitor polyiso-butylene （PiB） on their expressions and U87 cell proliferation.Methods Eighty-four glioma samples （15 with WHO graded Ⅱ,27 with WHO graded Ⅲ and 42 with WHO graded Ⅳ）,collected in our hospital from March 2013 to August 2014,were chosen in our study; their expressions of PIN1 and Nanog were detected by immunohistochemical staining; U87 glioblastoma cell lines were cultured; 0.2,0.5,1.0 and 2.0 μg/mL PiB was given to the cells for 24 and 48 h,and then,MTT assay was employed to detect the inhibited effect and inhibition ratio was calculated; 1.0 μmol/L PiB group （U87 cells treated with 1.0 μmol/L PiB for 24 h） and control group （U87 cells without any treatment） were chosen,immunofluorescence staining was used to detect the PIN1 and Nanog protein expressions,and real time-PCR and Western blotting were employed to observe the mRNA and protein expressions of PIN1 and Nanog.Results The positive protein expression rate of PIN1 and Nanog in gliomas of different grades （WHO graded Ⅱ,Ⅲ and Ⅳ） was significantly different （P〈0.05）：the higher the malignancy grade,the higher the rate of positive protein expression of PIN1 and Nanog.MTT assay indicated that PiB could inhibit the cell proliferation in dose-and concentration-dependent manners; as compared with the control group,1.0 μmol/L PiB group had significantly decreased percentages of PIN1＋,Nanog＋ and PIN1＋/Nanog＋ cells （P〈0.05）; the mRNA PIN1 expression quantity in the control group and 1.0 μmol/L PiB group was 1.82±0.03 and 0.94±0.20,and that of Nanog was 1.47±0.04 and 0.82±0.19,with significant differences （P〈0.05）; the protein PIN1 expression quantity in the control group and 1.0 μmol/L PiB group was 2.96±0.05 and 1.15±0.26,and that of Nanog was 1.52±0.16 and 0.75±0.05,with significant differences （P〈0.05）.Conclusion PIN1 and Nanog play important rol

关 键 词：神经胶质瘤 肽基脯氨酰异构酶1 NANOG 

分 类 号：R739.41[医药卫生—肿瘤]