COX-2抑制人急性髓系白血病OCI-AML2细胞株内p53、TNF-α的表达  

COX-2 inhibited the expressions of p53 and TNF-αin human acute myeloid leukemia OCI-AML2 cell line

作　　者：陈文婷[1] 林丽娥[1] 刘丹 黄莹 CHEN Wen-ting;LIN Li-e;LIU Dan;HUANG Ying(Hainan General Hospital,Haikou,Hainan 571100,China)

机构地区：[1]海南省人民医院血液内科,海南海口571100

出　　处：《热带医学杂志》2020年第7期882-886,906,共6页Journal of Tropical Medicine

基　　金：海南省科技计划项目(ZDYF2019186)。

摘　　要：目的探究抑制环氧化酶-2(COX-2)后对人急性髓系白血病OCI-AML2细胞增殖、凋亡的影响及其可能作用机制。方法体外培养人急性髓系白血病OCI-AML2细胞,细胞分为三组:空白对照组(不进行转染)、阴性对照(NC)组(转染阴性序列)、COX-2 RNAi组(转染COX-2 RNAi序列)。CCK-8法检测各组细胞增殖活性;流式细胞术检测转染后各组细胞凋亡、周期分布情况,免疫印迹法检测细胞中COX-2、Cyclin D1、p21、p53、肿瘤坏死因子-α(TNF-α)、Bax、Bcl-2蛋白表达。结果与空白对照组、阴性对照组比较,COX-2 RNAi组细胞COX-2蛋白表达(0.21±0.03)降低,细胞增殖抑制率(21.74±3.55)%、凋亡率(26.45±4.16)%升高,G1期细胞比例(44.17±1.36)%升高,S期(46.45±1.16)%、G2期(9.38±1.17)%细胞比例降低,p21(0.84±0.07)、p53(0.72±0.06)、Bax(1.26±0.15)蛋白表达升高,Cyclin D1(0.86±0.09)、TNF-α(0.45±0.08)、Bcl-2(0.24±0.03)蛋白表达降低,以上差异均有统计学意义(P<0.05)。结论抑制COX-2后能够抑制人急性髓系白血病OCI-AML2细胞增殖,促进其凋亡,使细胞周期阻滞于G1期,其机制可能是通过上调p53,下调TNF-α表达实现的。Objective To investigate the effects of cyclooxygenase-2(COX-2)inhibition on the proliferation and apoptosis of human acute myeloid leukemia OCI-AML2 cells,and explore the possible mechanism.Methods Human acute myeloid leukemia OCI-AML2 cells were cultured in vitro and divided into three groups:blank control group(no transfection).negative control(NC)group(transfection negative sequence),and COX-2 RNAi group(transfection COX-2 RNAi sequence).CCK-8 method was used to detect the proliferation activity of cells in each group;flow cytometry was used to detect the apoptosis and cycle distribution of cells in each group after transfection,and the expressions of COX-2,Cyclin D1,p21,p53,TNF-α,Bax and Bcl-2 were detected by Western blotting.Results Compared with the control group and the NC group,the expression of COX-2 protein in the COX-2 RNAi group was reduced(0.21±0.03),and the cell proliferation inhibition rate(21.74±3.55)%and the apoptosis rate(26.45±4.16)%were increased;the proportion of cells in G1 phase increased(44.17±1.36)%;the proportion of cells in S phase(46.45±1.16)%and G2 phase(9.38±1.17)%decreased;p21(0.84±0.07),p53(0.72±0.06),Bax(1.26±0.15)protein expression increased;Cyclin D1(0.86±0.09),TNF-α(0.45±0.08),and Bcl-2(0.24±0.03)protein expression decreased(P<0.05).Conclusions Inhibition of COX-2 could inhibit the proliferation and promote apoptosis of human acute myeloid leukemia OCI-AML2 cells,and block the cell cycle in G1 phase.The mechanism might be related to up-regulation of p53 and down-regulation of TNF-αexpression.

关 键 词：人急性髓系白血病 环氧化酶-2 肿瘤坏死因子-α P53 

分 类 号：R733.7[医药卫生—肿瘤]