高浓度胰岛素对K562细胞株增殖的抑制作用及机理  

Inhibitory Effect of High Concentration Insulin on K562 Cell Proliferation and Its Mechanism

作　　者：黄月琴[1] 潘宏达[2] 郭奕斌[3] 潘敬新[1] 

机构地区：[1]福建医科大学附属二院血液科,福建泉州362000 [2]北京大学肿瘤医院北京肿瘤医院结直肠外科,北京100142 [3]中山大学中山医学院遗传教研室,广东广州510080

出　　处：《中国实验血液学杂志》2016年第2期411-415,共5页Journal of Experimental Hematology

摘　　要：目的:探讨高浓度胰岛素对K562细胞株增殖抑制作用及其机理。方法:采用不同浓度胰岛素和/或抗IGF-1R抗体(IGF-1R-Ab)分别处理K562细胞,应用M TT法检测K562细胞的增殖活性,流式细胞术检测K562细胞凋亡的变化,Western blot法测定不同浓度胰岛素作用下K562细胞的IGF-1R、Akt、Erk1/2蛋白表达及磷酸化水平变化。结果:MTT法显示,低于40 mU/ml胰岛素具有促进K562细胞增殖,而高浓度(>40 m U/ml)胰岛素却显示出抑制K562细胞增殖活性。流式细胞术结果显示,40 mU/ml胰岛素抑制K562细胞凋亡(P<0.05),而200m U/ml胰岛素明显诱导K562细胞凋亡(P<0.01);0.01μg/ml至1.0μg/ml浓度IGF-1R-Ab具有显著增强高浓度(>40 m U/ml)胰岛素抑制K562细胞增殖和诱导K562细胞凋亡的作用(r=0.962,P<0.001)。Western blot结果显示,不同浓度胰岛素作用K562细胞后,ERK及p-ERK表达变化不明显,200 mU/ml浓度胰岛素作用K562细胞后,IGF-1R和AKT表达变化不明显,而IGF-1R和AKT磷酸化水平增强。结论:高浓度(>40 mU/ml)胰岛素能抑制K562细胞增殖并诱导其凋亡,其作用机制可能与胰岛素结合IGF-1R,竞争性地抑制IGF-1与IGF-1R的结合,相对阻断PI3K/AKT信号通路传导有关。Objective:To investigate the inhibitory effect of high concentration insulin on K562 cell proliferation and its underlying mechanism.Methods:K562 cells were treated by different concentration of insulin and / or anti-IGF-1R antibody(IGF-1R-Ab),MTT assay and flow cytometry were used to detect the K562 cells proliferation and apoptosis,respectively;Western blot was used to measure the expression and phosphorylation level of IGE-IR,Akt,Erk1 / 2 in K562 cells under the different concentration of insulin.Results:MTT assay showed that less than 40 mU / ml insulin could promote K562 cell proliferation,while high concentration(> 40 mU/ml) insulin has been shown to inhibit K562 cell proliferation;Flow cytometry showed that 40 mU /ml insulin suppressed K562 cell apoptosis(P < 0.05),while 200mil/ml insulin could significantly induce K562 cell apoptosis(P < 0.01);0.01 to 1.0 μg/ml IGF-1R-Ab has significantly enhanced the inhibitory and inducing effects of high concentration(> 40 mU/ml) of insulin on K562 cell proliferation and apoptosis respectively(r =0.962,P <0.001);Western blot showed that after K562 cells were treated with different concentrations of insulin ERK,and the p-ERK expression did not change significantly,after K562 cells were treated with 200 mU/ml insulin,the expression of IGF-1R and AKT also not were changed obviously,while the phosphorylation level of IGF-1R and AKT increased.Conclusion:High concentration(> 40 mU/ml) of insulin inhibits K562 cell proliferation and induces its apoptosis,and its mechanism may be related with the binding IGF-1R by insulin,competitively inhibiting the binding of IGF-1 and IGF-1R,the blocking the transduction of PI3 K/AKT signal pathway.

关 键 词：胰岛素 胰岛素生长因子1受体-抗体 K562细胞 细胞增殖 信号通路 

分 类 号：R73[医药卫生—肿瘤]