Par-6／aPKC极性蛋白重排促进胶质瘤侧群细胞快速增殖与分化  

Par -6/aPKC polarity protein rearrangement promoted cell rapid proliferation and differentiation of glioma side population cells

作　　者：刘晓智[1] 姜忠敏[2] 于士柱[4] 黄建永[5] 怀鹏[1] 李艳霞[3] 刘振林[1] 

机构地区：[1]天津市第五中心医院神经外科,300450 [2]天津市第五中心医院病理科,300450 [3]天津市第五中心医院中心实验室,300450 [4]天津医科大学总医院神经病学研究所 [5]杜克大学生物医学工程系,美国北卡罗来纳州

出　　处：《中华神经外科杂志》2014年第7期729-733,共5页Chinese Journal of Neurosurgery

基　　金：基金项目：国家自然科学基金项目（81000901）;天津市卫生局科技基金项目（2011KZ26和2011KZ24）;滨海新区医药卫生科技项目（2011BHKZ004、2012BWKL001和2012BWKZ001）

摘　　要：目的 探讨Par-6/非典型蛋白激酶C（atypical protein kinase C,aPKC）极性蛋白重分布是否能促进胶质瘤侧群细胞快速增殖与分化.方法 实验分为对照组、无义序列组和siR-PKCξ组3组.免疫细胞荧光方法检测aPKC、Par-6和TRIM32（tripartite motif-containing protein 32 gene）在细胞中的定位,绘制细胞球体生长曲线,Ki67免疫荧光方法检测细胞核增殖活性,免疫荧光方法检测细胞标记蛋白的表达.结果 与对照组和无义序列组比较,siR-PKCξ组PKCζ蛋白表达水平明显降低（0.07±0.03,P＜0.01）,Par-6极性消失,TRIM32发生核转移.基因转染24 h,siR-PKCξ组Ki67阳性百分率为（48.65±11.34）％,均高于其他两组（P＜0.05）,转染72 h,siR-PKCζ组低表达CD133和Nestin,高表达GFAP（P ＜0.05）,β-tubulinⅢ和myelin表达无明显变化（P＞0.05）.结论 通过诱导Par-6/aPKC极性蛋白重排可促进胶质瘤侧群细胞快速增殖与分化.Objective To promote cell rapid proliferation and differentiation of glioma side population cells by inducing redistribution of Par-6/atypical protein kinase C （aPKC） polarity proteins.Methods The experiments was divided into control group,non-antisense group and siR-PKCξ group.The cellular immunofluorescence staining was used to detect the distribution of aPKC,Par-6 and tripartite motif-containing protein 32 gene （TRIM32） in cells.The growth curves were drawn,and the nuclear proliferation was detected by immunofluorescence for Ki67.The expression of cell markers was detected by immunofluorescence method.Results Compared with the control group and non-antisense group,the expression level of siR-PKCξ in siR-PKCξ group were significantly lower （0.07-± 0.03,P 〈 0.01）,accompanied by loss of Par-6 polarity and TRIM32 nuclear transfer.At 24h after gene transfection,Ki67 positive percentage was （48.65 ± 11.34）%,which was higher than those in the other two groups （P 〈 0.05）.At 72h after gene transfection,there were lower expression of CD133 and nestin,and higher expression of GFAP （P 〈 0.05）,but there was no significant changes of 3-tubulin Ⅲ and myelin in siR -PKCξ group （P 〉 0.05）.Conclusion Rearrangement of Par-6/aPKC polarity protein could promote rapid proliferation and differentiation of glioma side population cells.

关 键 词：神经胶质瘤 细胞极性 RNA干扰 细胞周期 

分 类 号：R739.41[医药卫生—肿瘤]