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目的:观察相对分子质量小于10000的小分子胸腺肽(thymopeptide,TP)在体外液体培养中对人急性早幼粒细胞白血病细胞系HL60细胞增殖的影响。并探讨可能的作用机制。方法:在加或不加TP的情况下,体外悬浮培养HL60细胞。实验分为3组:空白对照组、TP20μg/ml和40μg/ml实验组,每组设3个平行孔;胎盘蓝染色法计数细胞并绘制细胞生长指数曲线;单细胞凝胶电泳法(SCGE)检测细胞出现的DNA损伤;琼脂糖凝胶电泳法检测基因组DNA断裂情况。结果:TP作用于细胞72h后,较低浓度的TP(20μg/ml)对HL60细胞的增殖即有抑制作用;对TP两个实验组的SCGE检测结果显示,HL60细胞的慧尾现象明显增多;琼脂糖凝胶电泳上出现了相差200bp左右的DNA梯度现象。结论:TP对HL60细胞的增殖具有抑制作用,凋亡诱发的DNA损伤可能参与了这一过程。
OBJECTIVE: To observe the effect of thymopeptide (TP) with molecular weight less than 10000 on the proliferation of human acute promyelocytic leukemia HL60 cells in vitro. And explore the possible mechanism of action. Methods: HL60 cells were cultured in vitro with or without TP. The experiment was divided into three groups: blank control group, TP20μg / ml and 40μg / ml experimental group, with 3 parallel wells in each group; cells were counted by placental staining and cell growth index curve was drawn; single cell gel electrophoresis (SCGE) Detection of cell DNA damage occurs; agarose gel electrophoresis assay genomic DNA fragmentation. Results: After treated with TP for 72h, the proliferation of HL60 cells was inhibited by lower concentration of TP (20μg / ml). The results of SCGE in two experimental groups showed that the homing of HL60 cells was significantly increased. Agarose gel electrophoresis appeared a difference of about 200bp DNA gradient phenomenon. Conclusion: TP can inhibit the proliferation of HL60 cells, and DNA damage induced by apoptosis may be involved in this process.