上海交通大学学报（医学版）
上海交通大學學報（醫學版）
상해교통대학학보（의학판）
JOURNAL OF SHANGHAI JIAOTONG UNIVERSITY(MEDICAL SCIENCE)
2010年
2期
125-128,139
,共5页
林芊%吴伟岚%卫敏江%沈加%谈珍%施君%董华腾%李玉峰
林芊%吳偉嵐%衛敏江%瀋加%談珍%施君%董華騰%李玉峰
림천%오위람%위민강%침가%담진%시군%동화등%리옥봉
细胞凋亡%caspase-3%细胞周期%Fas
細胞凋亡%caspase-3%細胞週期%Fas
세포조망%caspase-3%세포주기%Fas
apoptosis%caspase-3%cell cycle%Fas
目的 研究Fas诱导的B淋巴细胞瘤细胞株MML-1凋亡过程中,细胞内活化caspase-3表达与细胞周期的关系.方法 抗Fas抗体孵育MML-1不同时间以诱导细胞凋亡,流式细胞仪检测凋亡率并验证MML-1对Fas诱导凋亡的敏感性.采用PI-Triton X和FITC-active caspase-3双染色分析经Fas诱导凋亡的MML-1内活化caspase-3的表达.以细胞周期蛋白cyclin A、B_1、E 标记Fas诱导凋亡的处于不同增殖周期MML-1(S、G_2/M和G_1期细胞),流式细胞术检测细胞内活化caspase-3的表达.结果 Fas诱导6 h后,MML-1凋亡率达到56%.Fas诱导4 h后,活化caspase-3在G_1期细胞内表达显著升高,仅有少量S期和G_2/M期细胞表达活化caspase-3.Cyclin A、B_1阴性而cyclin E阳性MML-1表达活化caspase-3.结论 Fas诱导MML-1凋亡过程中,细胞内活化caspase-3的表达与细胞周期有关.G_1后期和S早期的MML-1最先表达活化caspase-3,且对Fas诱导凋亡的敏感性最高.
目的 研究Fas誘導的B淋巴細胞瘤細胞株MML-1凋亡過程中,細胞內活化caspase-3錶達與細胞週期的關繫.方法 抗Fas抗體孵育MML-1不同時間以誘導細胞凋亡,流式細胞儀檢測凋亡率併驗證MML-1對Fas誘導凋亡的敏感性.採用PI-Triton X和FITC-active caspase-3雙染色分析經Fas誘導凋亡的MML-1內活化caspase-3的錶達.以細胞週期蛋白cyclin A、B_1、E 標記Fas誘導凋亡的處于不同增殖週期MML-1(S、G_2/M和G_1期細胞),流式細胞術檢測細胞內活化caspase-3的錶達.結果 Fas誘導6 h後,MML-1凋亡率達到56%.Fas誘導4 h後,活化caspase-3在G_1期細胞內錶達顯著升高,僅有少量S期和G_2/M期細胞錶達活化caspase-3.Cyclin A、B_1陰性而cyclin E暘性MML-1錶達活化caspase-3.結論 Fas誘導MML-1凋亡過程中,細胞內活化caspase-3的錶達與細胞週期有關.G_1後期和S早期的MML-1最先錶達活化caspase-3,且對Fas誘導凋亡的敏感性最高.
목적 연구Fas유도적B림파세포류세포주MML-1조망과정중,세포내활화caspase-3표체여세포주기적관계.방법 항Fas항체부육MML-1불동시간이유도세포조망,류식세포의검측조망솔병험증MML-1대Fas유도조망적민감성.채용PI-Triton X화FITC-active caspase-3쌍염색분석경Fas유도조망적MML-1내활화caspase-3적표체.이세포주기단백cyclin A、B_1、E 표기Fas유도조망적처우불동증식주기MML-1(S、G_2/M화G_1기세포),류식세포술검측세포내활화caspase-3적표체.결과 Fas유도6 h후,MML-1조망솔체도56%.Fas유도4 h후,활화caspase-3재G_1기세포내표체현저승고,부유소량S기화G_2/M기세포표체활화caspase-3.Cyclin A、B_1음성이cyclin E양성MML-1표체활화caspase-3.결론 Fas유도MML-1조망과정중,세포내활화caspase-3적표체여세포주기유관.G_1후기화S조기적MML-1최선표체활화caspase-3,차대Fas유도조망적민감성최고.
Objective To investigate the relationship between expression of active form of caspase-3 and cell cycle in Fas-mediated apoptosis of B lymphocytoma cell line MML-1. Methods MML-1 cells were incubated with agonistic anti-Fas antibody for different time,and cell apoptosis was induced.Cell apoptotic rates were analysed by flow cytometry,and sensitivity of MML-1 cells to apoptosis was determined.The expression of active form of caspase-3 was analysed by double staining with PI-Triton X and FITC-active caspase-3.Cyclin A,B_1 and E were selected as cell cycle markers for S,G_2/M and G_1 phase of MML-1 cells,and the expression of active form of caspase-3 was detected by flow cytometry. Results The cell apoptotic rate reached 56% after induction by Fas for 6 h.After induction by Fas for 4 h,the active form of caspase-3 was mainly expressed in cells of G_1 phase,while rarely in cells of S and G_2/M phase.Cells with negative cyclin A and B_1 and positive cyclin E expressed active form of caspase-3. Conclusion The expression of active form of caspase-3 in MML-1 cells mediated by Fas might be cell cycle dependent.Cells entering into late G_1 and early S phase first express active form of caspase-3,and their sensitivity to Fas-mediated apoptosis is the highest.
