CAJ | 학술논문

目的有丝分裂纺锤体和中间小体均是以在细胞有丝分裂进程中出现的微管为结构基础的暂时性结构,对细胞有丝分裂的顺利进行起着至关重要的作用.本研究旨在建立成熟高效的提取细胞内有丝分裂纺锤体和中间小体的方法.方法通过细胞周期同步化方法使细胞内出现有丝分裂纺锤体或中间小体,运用低渗肿胀和甘油梯度离心的原理提取纺锤体和中间小体.结果经Western Blot和细胞免疫荧光染色法鉴定,证实提取物中含有有丝分裂纺锤体和中间小体.结论建立从人宫颈癌细胞Hela中提取有丝分裂纺锤体及中间小体的方法,可为鉴定有丝分裂纺锤体及中间小体上的蛋白分子垫定实验基础,同时也对研究肿瘤细胞有丝分裂的分子调控机制具有重要意义.
목적 유사분렬방추체화중간소체균시이재세포유사분렬진정중출현적미관위결구기출적잠시성결구,대세포유사분렬적순리진행기착지관중요적작용.본연구지재건립성숙고효적제취세포내유사분렬방추체화중간소체적방법.방법 통과세포주기동보화방법사세포내출현유사분렬방추체혹중간소체,운용저삼종창화감유제도리심적원리제취방추체화중간소체.결과 경Western Blot화세포면역형광염색법감정,증실제취물중함유유사분렬방추체화중간소체.결론 건립종인궁경암세포Hela중제취유사분렬방추체급중간소체적방법,가위감정유사분렬방추체급중간소체상적단백분자점정실험기출,동시야대연구종류세포유사분렬적분자조공궤제구유중요의의.
Objective Mitotic spindle and midbody are both microtubule-based temporary structures during cell growth and play essential roles in mitosis.The purpose of this study was to establish a mature and efficient method to extract mitotic spindle and midbody.Methods Through the cell cycle synchronization method,mitotic spindle or midbody was made appear inside cells.Low permeability swelling and glycerol gradient centrifugation principles were then used to extract spindle and midbody.Results By Western Blot and immunofluorescence staining,the extracts were identified as mitotic spindle and midbody.Conclusions The successful extraction of mitotic spindle and midbody from synchronized Hela cells will provide foundation for identifying the proteins located in cell during mitosis,and be of great significance to the study of molecular regulation mechanisms of mitosis and tumorigenesis.