目的:探讨胶质瘤细胞中p27酬蛋白的表达、定位,为进一步研究p27kip1在胶质瘤发生、发展过程中的功能奠定理论基础。方法:用免疫荧光方法检测U87、LN308细胞p27kiP1蛋白的定位情况;进一步分离两种细胞的细胞质与细胞核,在显微镜下观察细胞核形态并用DAPI染色分析细胞核完整性,提取蛋白用Westernblotting。方法检测分离的细胞质与细胞核蛋白的纯度,并检测p27kip1,在细胞中的表达情况。结果:成功分离了细胞的细胞浆与细胞核,并得到纯度较好的细胞浆蛋白与细胞核蛋白。确定了p27kip1蛋白主要表达于U87和LN308细胞的细胞质中。结论:p27kip1蛋白在恶性胶质瘤中可能主要表达在细胞质中,并且其亚细胞定位可能与胶质瘤的恶性程度相关。
Objective: To investigate the expression and localization of p27kiP1; in glioma cells, and give the basis function for analysis of p27kiP1; in the tumorigenesis and development of glioma. Methods: The localization of p27kiP1; in U87 cells and LN308 cells was detected by immunofluorescence. The cytoplasm and nucleus was seperated, and the nucleus were stain with DAPI. Then the integrity of nucleu was detected under a microscope. The purity of the protein and the expression of p27kiP1; in the cytoplasm and nucleus were detected by using western blotting assay. Results: The cytoplasm and nucleus were seperated, and it was found that the p27kiP1 protein expressed mainly in the cytoplasm of U87 cells and LN308 ceUs. Conclusion: The p27kiP1 protein expressed mainly in the cytoplasm of glioma, and the subcellular localization of p27kiP1 may be consistent with their biological ftmction.