海生素对K562细胞生长及凋亡基因表达的影响

目的 探讨海生素对白血病K562细胞生长及凋亡基因表达的影响. 方法实验分为对照组、10mg/L海生素组和20rag/L海生素组.采用流式细胞术(FcM)、四唑蓝(MTT)和免疫细胞化学法测定海生素对K562细胞周期、细胞生长及细胞凋亡基因bcl-2和bax表达的影响. 结果海生素浓度为10mg/L和20mg/L时,可阻止K562细胞周期于G1/S期;海生素浓度为20mg/L时对K562细胞有抑制作用,在此浓度下随时间的延长细胞存活率有下降趋势;海生素浓度为20mg/L和40mg,L时,可下调凋亡抑制基因bcl-2的表达,上调凋亡促进基因hax的表达.结论 海生素可明显抑制K562细胞的增殖,使其阻滞于细胞周期的G1/S期;海生素还通过减少bcl-2的表达及增加bax的表达从而促进K562细胞的凋亡.

INFLUENCE OF HAISHENGSU ON CELL GROWTH AND EXPRESSION OF APOPTOSIS GENE IN K562 CELLS

Objective To study the influence of Haishengsu on cell growth and the expression of apoptosis gene in leukemia K562 cells by Haishengsu. Methods K562 cells were cultured in RPMI1640 containing 10% fetal bovine serum at 37℃ in a humidified atmosphere with 5% CO2. The experiment was performed on three groups: the control group and two Haishengsu groups (10mg/L and 20mg/L).The cell cycle, growth and the expression of Bcl-2 and bax were evaluated respectively by means of flow cytometer(FCM), thiazoyl blue tetrazolium bromide(MTT)assay and immunocytochemistry to determine Haishengsu’s influence. Results Both the Haishengsu (10mg/L and 20mg/L), restrained cell cycle in G1/S phase. The 20mg/L inhibited the growth of K562 cells and the survival of cells declined along with the time in this concentration. It was observed under light microscope that the number of positive cells increased in the two Haishengsu groups(20mg/L and 40mg/L).It was analyzed by image analysis instrument that the expression of Bcl-2 proportion decreased to (58.78±4.65)% and (26.57±2.13)% respectively in the Haishengsu groups (20mg/L and 40mg/L) from (90.98±8.66)% in the control group and the expression of bax proportion increased to (77.69±3.55)% and (90.60±3.73)% respectively in the Haishengsu groups (20mg/L and 40mg/L) from (10.88±6.57)% in the control group.Haishengsu (20mg/L and 40mg/L) could down regulate the expression of Bcl-2 and up regulate the expression of bax in K562 cells. Conclusion Haishengsu could significally restrain the cell cycle in G1/S phase and inhibit the growth of K562 cells. Haishengsu could reduce the expression of Bcl-2 and increase th