Erbin基因对急性髓系白血病细胞周期、增殖、凋亡和分化的影响  

Effect of Erbin on proliferation and differentiation of acute myeloid leukemia cells

作　　者：郑卓军[1] 顾伟英[1] 谢晓宝[1] 蒋敬庭[1] 

机构地区：[1]苏州大学附属第三医院肿瘤中心 苏州大学细胞治疗研究院,常州213003

出　　处：《中华实验外科杂志》2018年第2期319-322,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金（31570877、31570908）;国家科技支撑计划（2015BA112812）

摘　　要：目的探讨Erbin与急性髓系白血病（AML）细胞周期、增殖、凋亡及分化的关系。方法通过实时定量聚合酶链反应（Real-time PCR）和Western blot检测5种AML细胞株HL60、THP-1、SHI-1、U937和NB4中内源性Erbin表达，并挑选出相对表达高及表达低的细胞各1株。转染Erbin过表达质粒慢病毒使其在低表达细胞株中过表达，转染短发卡RNA慢病毒使Erbin在高表达细胞株中下调。采用细胞计数试剂盒（CCK-8）及流式细胞术检测上述两种细胞和各自空载体对照组细胞周期、增殖、凋亡及分化情况，以及与细胞周期调节蛋白p21Waf1/CIP1、p27Kip1表达的关系。结果U937中Erbin相对表达量较高，为2.64±0.05，HL60和SHI-1细胞中表达量较低，为1.02±0.05及0.83±0.05。过表达Erbin的HL60细胞48 h细胞活性为0.65±0.02，较对照组（0.78±0.02）显著降低（t＝7.961，P＝0.001），G2/M期细胞比例[（26.00±0.44）%]较对照组[（33.24±1.29）%]降低（t＝5.311，P＝0.006），凋亡率[（9.51±0.81）%]较对照组[（2.47±1.13）%]增高（t＝8.765，P＝0.000），细胞周期调节蛋白p21Waf1/CIP1、p27Kip1表达升高，分化程度[CD11b表达率：（53.02±4.51）%]较对照组[CD11b表达率：（38.59±4.62）%]升高（t＝3.870，P＝0.018）。下调Erbin的U937细胞48 h细胞活性为0.68±0.04，较对照组（0.56±0.05）显著增高（t＝3.246，P＝0.031），G2/M期细胞比例[（20.30±0.83）%]较对照组[（14.76±0.86）%]升高（t＝4.635，P＝0.009），p21Waf1/CIP1、p27Kip1表达降低，分化程度[CD11b表达率：（30.52±3.57）%]较对照组[CD11b表达率：（51.26±6.21）%]下降（t＝5.019，P＝0.007）。结论Erbin在AML中通过抑制细胞增殖、细胞周期，促进细胞凋亡及细胞分化来发挥抗肿瘤活性。Objective To elucidate the impact of Erbin on the cell cycle, proliferation, apoptosis and differentiation of acute myeloid leukemia （AML） cells.Methods The expression levels of Erbin in acute myeloid 1eukemia cell lines （HL60, THP-1, SHI-1, U937 and NB4） were analyzed by real-time quantitative polymerase chain reaction （Real-time PCR） and Western blotting. We exogenously increased the level of Erbin in the leukemia cell line with low-Erbin-expression and exogenously reduced the level of Erbin in the leukemia cell line with high-Erbin-expression. The proliferation capacity was assayed by cell counting kit-8 （CCK-8）, cell cycle by Hoechst/PY, and apoptosis by Annexin V and differentiation by flow cytometry.Results U937 was selected as cell line with high-Erbin-expression and HL60 was selected as cell line with low-Erbin-expression. We found that proliferation （0.65±0.02 vs. 0.78±0.02） and G2/M cell cycle [（26.00±0.44）% vs. （33.24±1.29）%] were inhabited, early apoptosis [（9.51±0.81）% vs. （2.47±1.13）%] and differentiation were promoted [CD11b： （53.02±4.51）% vs. （38.59±4.62）%] in HL60 with exogenously elevated level of Erbin. Theaccordant results were also observed in U937 with exogenously decreased level of Erbin. The proliferation （0.68±0.04 vs. 0.56±0.05） and G2/M cell cycle [（20.30±0.83）% vs. （14.76±0.86）%] were promoted, differentiation [CD11b： （30.52±3.57）% vs. （51.26±6.21）%] was inhabited. Besides, the expression of Erbin was positively correlated with the expression of p21Waf1/CIP1 and p27Kip1.Conclusion Erbin may act as a cancer suppressor gene in AML cells, and may be a novel target for AML.

关 键 词：急性髓系白血病 Erbin基因 细胞增殖 细胞凋亡 分化 

分 类 号：R733.71[医药卫生—肿瘤]