CAJ | 학술논문

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.
목적 탐토Par-6/비전형단백격매C(atypical protein kinase C,aPKC)겁성단백중분포시부능촉진효질류측군세포쾌속증식여분화.방법 실험분위대조조、무의서렬조화siR-PKCξ조3조.면역세포형광방법검측aPKC、Par-6화TRIM32(tripartite motif-containing protein 32 gene)재세포중적정위,회제세포구체생장곡선,Ki67면역형광방법검측세포핵증식활성,면역형광방법검측세포표기단백적표체.결과 여대조조화무의서렬조비교,siR-PKCξ조PKCζ단백표체수평명현강저(0.07±0.03,P＜0.01),Par-6겁성소실,TRIM32발생핵전이.기인전염24 h,siR-PKCξ조Ki67양성백분솔위(48.65±11.34)％,균고우기타량조(P＜0.05),전염72 h,siR-PKCζ조저표체CD133화Nestin,고표체GFAP(P ＜0.05),β-tubulinⅢ화myelin표체무명현변화(P＞0.05).결론 통과유도Par-6/aPKC겁성단백중배가촉진효질류측군세포쾌속증식여분화.
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.