论文部分内容阅读
目的探讨高浓度胰岛素(insulin,INS)对K562细胞株增殖的抑制作用及其机理。方法用不同浓度胰岛素处理K562细胞,分别用CCK-8法、细胞计数法和台盼蓝拒染法检测K562细胞的增殖活性,同时检测培养基中葡萄糖浓度变化;流式细胞术检测K562细胞的凋亡效应;并分别采用不同浓度的胰岛素样生长因子-1(IGF-1)和IGF-1受体非特异性阻滞剂苏拉明(Suramin)对高浓度胰岛素影响K562细胞增殖作用进行干预。结果胰岛素在0.1~1mU/mL浓度下具有明显促进K562细胞增殖的作用,而高浓度胰岛素(1.6~100mU/mL)则相反;无论培养基中葡萄糖浓度高低,高浓度胰岛素都有对K562细胞增殖的抑制作用;高浓度胰岛素抑制K562细胞增殖呈时效和量效关系;0.1、1、10、100mU/mL胰岛素处理K562细胞48h,与对照组比较,0.1、1mU/mL浓度下抑制细胞凋亡,而10、100mU/mL浓度下促进细胞凋亡(P均<0.05);IGF-1能逆转高浓度胰岛素对K562细胞增殖的抑制作用,并且具有量效和时效关系;Suramin能增强高浓度胰岛素对K562细胞增殖的抑制作用,并且具有量效和时效关系。结论胰岛素对K562细胞有双重作用,即在0.1~1 U/mL浓度下促进细胞增殖、抑制凋亡,高浓度(1.6~100mU/mL)下抑制细胞增殖、促进凋亡,且其抑制作用与培养基中葡萄糖代谢无关与抑制IGF-1途径有关。
Objective To investigate the inhibitory effect of insulin (INS) on the proliferation of K562 cell line and its mechanism. Methods The K562 cells were treated with different concentrations of insulin, and the proliferation activity of K562 cells was detected by CCK-8 assay, cell counting method and trypan blue exclusion assay, respectively. Meanwhile, the glucose concentration in culture medium was also measured. Flow cytometry The apoptosis of K562 cells was induced by high concentrations of insulin using different concentrations of insulin-like growth factor-1 (IGF-1) and IGF-1 receptor nonspecific blocker Suramin. Results Insulin could significantly promote the proliferation of K562 cells at a concentration of 0.1 ~ 1 mU / mL, while high concentration of insulin (1.6 ~ 100 mU / mL) reversed the effect of insulin on proliferation of K562 cells regardless of glucose concentration K562 cells treated with 0.1, 1, 10 and 100 mU / mL insulin for 48h, compared with the control group, 0.1,1mU / mL inhibited the apoptosis of K562 cells, (P <0.05). IGF-1 could reverse the inhibitory effect of high concentrations of insulin on the proliferation of K562 cells with dose-effect and time-dependent effects. Suramin could enhance the effect of high concentration of insulin on the proliferation of K562 cells K562 cell proliferation, and has a dose-effect and time-dependent relationship. Conclusion Insulin has a dual effect on K562 cells, that is, it can promote cell proliferation and inhibit apoptosis at a concentration of 0.1-1 U / mL, and inhibit proliferation and apoptosis at high concentration (1.6-100 mU / mL) Glucose metabolism in culture medium has nothing to do with the inhibition of IGF-1 pathway.