利用RNA干涉文库进行大规模高通量的功能基因扫描,已成为发现新功能基因的重要方式和手段.为了寻找在细胞增殖和分化过程中的新功能基因,根据斯坦福大学公布的与人类胚胎干细胞和造血干细胞增殖和分化过程中有关基因的基因芯片的分析结...利用RNA干涉文库进行大规模高通量的功能基因扫描,已成为发现新功能基因的重要方式和手段.为了寻找在细胞增殖和分化过程中的新功能基因,根据斯坦福大学公布的与人类胚胎干细胞和造血干细胞增殖和分化过程中有关基因的基因芯片的分析结果,组建了与细胞增殖和分化有关的RNA限制性干涉文库.该文库包括靶向各类基因的载体,如包括转录因子、各类蛋白激酶、细胞周期调控蛋白以及一些未知功能基因在内的225个基因.利用这个限制性RNA干涉文库对控制HeLa细胞增殖的基因进行筛选.并通过WST-1高通量检测,发现了2个同HeLa细胞增殖相关的基因:CNKSR3(Homo sapiens CNKSR family member 3)和Fosl2(Homo sapiens FOS-like antigen 2),并初步证实:沉默CNKSR3会促进HeLa细胞的增殖,而沉默Fosl2则抑制HeLa细胞的增殖功能.展开更多
To explore the role of a novel Obg-like ATPase 1 （OLA1） in cancer metastasis, small interference RNA （siRNA） was used to knockdown the protein, and the cells were subjected to in vitro cell migration and invasion ...To explore the role of a novel Obg-like ATPase 1 （OLA1） in cancer metastasis, small interference RNA （siRNA） was used to knockdown the protein, and the cells were subjected to in vitro cell migration and invasion assays. Knockdown of OLA 1 significantly inhibited cell migration and invasion in breast cancer cell line MDA-MB-231. The knockdown caused no changes in cell growth but affected ROS production. In wound-healing assays, decreased ROS in OLA1-knockdown cells were in situ asso- ciated with the cells＇ decreased motile morphology. Further, treatment ofN-acetylcysteine, a general ROS scavenger, blunted the motility and invasiveness of MDA-MB-231 cells, similar to the effect of OLAl-knockdown. These results suggest that knock- down of OLA1 inhibits breast cancer cell migration and invasion through a mechanism that involves the modulation of intracellular ROS levels.展开更多
文摘利用RNA干涉文库进行大规模高通量的功能基因扫描,已成为发现新功能基因的重要方式和手段.为了寻找在细胞增殖和分化过程中的新功能基因,根据斯坦福大学公布的与人类胚胎干细胞和造血干细胞增殖和分化过程中有关基因的基因芯片的分析结果,组建了与细胞增殖和分化有关的RNA限制性干涉文库.该文库包括靶向各类基因的载体,如包括转录因子、各类蛋白激酶、细胞周期调控蛋白以及一些未知功能基因在内的225个基因.利用这个限制性RNA干涉文库对控制HeLa细胞增殖的基因进行筛选.并通过WST-1高通量检测,发现了2个同HeLa细胞增殖相关的基因:CNKSR3(Homo sapiens CNKSR family member 3)和Fosl2(Homo sapiens FOS-like antigen 2),并初步证实:沉默CNKSR3会促进HeLa细胞的增殖,而沉默Fosl2则抑制HeLa细胞的增殖功能.
基金Project supported by the National Basic Research Program (973) of China (No. 2004CB518707) the Methodist Hospital Research Institute, USA
文摘To explore the role of a novel Obg-like ATPase 1 （OLA1） in cancer metastasis, small interference RNA （siRNA） was used to knockdown the protein, and the cells were subjected to in vitro cell migration and invasion assays. Knockdown of OLA 1 significantly inhibited cell migration and invasion in breast cancer cell line MDA-MB-231. The knockdown caused no changes in cell growth but affected ROS production. In wound-healing assays, decreased ROS in OLA1-knockdown cells were in situ asso- ciated with the cells＇ decreased motile morphology. Further, treatment ofN-acetylcysteine, a general ROS scavenger, blunted the motility and invasiveness of MDA-MB-231 cells, similar to the effect of OLAl-knockdown. These results suggest that knock- down of OLA1 inhibits breast cancer cell migration and invasion through a mechanism that involves the modulation of intracellular ROS levels.
