HMBA诱导人成骨肉瘤MG-63细胞分化过程中hnRNP A2/B1的表达与定位  

Expression and Localization of hnRNP A2/B1 during Differentiation of Human Osteosarcoma MG-63 Cells Induced by HMBA

作　　者：赵春红[1] 李祺福[1] 陈兰英[2] 唐剑[1] 宋建晔[1] 谢忠[1] 

机构地区：[1]厦门大学细胞生物学与肿瘤细胞工程教育部重点实验室,福建厦门361005 [2]平顶山工学院生物工程系,河南平顶山467001

出　　处：《癌症》2008年第7期677-684,共8页Chinese Journal of Cancer

基　　金：国家自然科学基金项目(No.30470877)~~

摘　　要：背景与目的:核基质蛋白的差异表达与细胞癌变和增殖分化调控关系密切。本研究观察了hnRNP A2/B1在诱导分化处理前后人成骨肉瘤MG-63细胞核基质中的存在和分布,及其与Actin、Prohibitin的共定位关系。方法:选择性抽提经环六亚甲基双乙酰胺(hexamethylene bisacetamide,HMBA)诱导处理前后的MG-63细胞核基质,并运用双向电泳、质谱分析、蛋白质印记杂交、免疫荧光、激光共聚焦等技术检测hnRNP A2/B1在核基质中的表达与定位变化,及其与相关蛋白的共定位关系。结果:双向电泳及蛋白质印迹杂交结果证实了hnRNP A2/B1存在于MG-63细胞核基质蛋白组分中,并在HMBA处理后细胞核基质中表达下调;免疫荧光显微镜观察显示hnRNP A2/B1定位在核基质上,经HMBA处理后hnRNP A2/B1表达减弱。激光共聚焦显微镜观察结果显示,hnRNP A2/B1与细胞核基质蛋白组分Actin、细胞增殖相关调控因子Prohibitin具有共定位关系,但在诱导处理后细胞内的共定位关系减弱。结论:hnRNP A2/B1在MG-63细胞诱导分化过程中的表达分布,及其与Actin、Prohibitin的共定位关系的改变对MG-63细胞分化具有重要影响,值得进一步探索和研究。BACKGROUND ＆ OBJECTIVE： The differentially expressed nuclear matrix proteins have great effects on canceration and regulation of cell differentiation. This study was to explore the existence and distribution of ribonucleoprotein hnRNP A2/B1 in nuclear matrix and its co-localization with Actin and Prohibitin in human osteosarcoma MG-63 cells before and after hexamethylene bisacetamide （HMBA） treatment. METHODS： The nuclear matrix of MG-63 cells before and after treatment of HMBA were selectively extracted. The expression and localization of hnRNP A2/B1 in nuclear matrix were detected by 2-D PAGE, MALDI-TOF-MS, Western blot, and immunofiuorescent staining. The co-localization of hnRNP A2/B1 with Actin and Prohibitin was observed under laser scanning confocal microscope （LSCM）. RESULTS： hnRNP A2/B1 was detected in the component of nuclear matrix proteins of MG-63 cells by Western blot and immunogold staining and its expression was decreased after treatment of HMBA. hnRNP A2/B1 was located in the nuclear matrix, and its expression was weakened after HMBA treatment, hnRNP A2/B1 was co-localized with Actin or Prohibitin in MG-63 cells, while the co-localization relationship was weakened during differentiation of MG-63 cells. CONCLUSIONS- hnRNP A2/B1 is a kind of nuclear matrix protein, and localizes in the nuclear matrix. The distribution and expression of hnRNP A2/B1 and its co-localization with Actin and Prohibitin play important roles during the differentiation of MG-63 cells.

关 键 词：HNRNP A2/B1 核基质 成骨肉瘤 MG-63细胞 细胞分化 

分 类 号：R738.1[医药卫生—肿瘤]