【摘要】 目的探讨蟾蜍灵对胃癌MGC803细胞周期及周期蛋白的影响。方法采用台盼蓝拒染法测定细胞增殖活力;瑞士姬姆萨染色观察细胞形态学的变化;流式细胞仪检测细胞周期;免疫组织化学方法检测细胞周期相关蛋白p16、p21、pRb的表达。结果1)蟾蜍灵抑制胃癌MGC803细胞的增殖,24、48、72h的IC50分别为0.086、0.048和0.036μmol/L。2)蟾蜍灵在0.01～0.1μmol/L时作用24～72h,形态学上可见细胞体积增大,出现双核、多核细胞。3)流式细胞仪显示蟾蜍灵浓度为0.01～0.1μmol/L时可明显诱导细胞周期G2/M期阻滞。4)蟾蜍灵作用于胃癌MGC803细胞,观察到p16、p21、pRb蛋白的表达明显上调。结论蟾蜍灵引起胃癌MGC803细胞的周期阻滞可能与p16、p21、pRb蛋白的上调有关。更多还原

【Abstract】 OBJECTIVE:To investigate the effect on cell c yc le of MGC-803 cell line induced by bufalin, and the change of protein expressio n accompanied with the cell cycle arrest.METHODS:Cell viab ility was counted by trypan blue dye exclusion, The morphological changes were o bserved with light microscope dyed in Wright-Giemsa, the distribution of cell c ycles was analyzed with flow cytometry （FCM）, and the expressions of protein p16 ,p21,pRb was assayed by immunocytochemical staining.RESULTS: In gastric cell lines MGC-803,1)Bufalin inhibits cell growth, and 50% inhibit ory concentrations （IC_<sup>50 ） of the agent at 24 h,48 h and 72 h were 0.086、 0.048、0.036 μmol/L, respectively. 2)When induced by bufalin at concentration of 0.01-0.1 μmol/L,the change in morphology including bigger volume, two or more nucleus was observed. 3)The flow cytometry analysis showed that 0.01-0.1 μmol/L treatment by bufalin resulted in accumulation of cells in the G₂/M pha se.4)After treated with bufalin, the expressions of protein p16,p21 and pRb were up-regulated.CONCLUSIONS:Bufalin inhibits gastric cancer MGC-803 cell proliferation, arrests the cells at the G₂/M phage of the cell cycle. These results may be related to the up-regulation of the p16,p21,pRb lev els.[更多还原