IL-23诱导人外周血单个核细胞对MUTZ-1细胞增殖、凋亡及相关基因表达的影响

摘要

Objective To observe the effects of peripheral blood mononuclear cell (PBMNCs)induced by IL-23 on the proliferation,apoptosis and gene expression of MUTZ-1 cells-a human myelodysplastic syndrome (MDS) cell line. Methods The PBMNCs were isolated by ficoll density gradient centrifugation. The cells were divided into four groups:control group (without IL-23),three different concentration IL-23 groups (2,10,50ng/ ml),which were induced in vitro for 72 hours. Then PBMNCs were co-cultured with MUTZ-1 cells. MTT assay was used to detect the proliferation of MUTZ-1 cells. Flow cytometry was used to detect the cell cycle and cell apoptosis. Real time-PCR and Western-blot were used to detect the expression levels of Bax,Bcl-2,caspase-3 and survivin mRNA and protein. Results After MUTZ-1 cells were co-cultured with PBMNCs induced by 2,10,50ng/ ml IL-23,respectively,the cell proliferation rate of MUTZ-1 and cell proportion at phase S were significantly decreased,however,the cell apoptosis rate and cell proportion at phase G0 / G1 were significantly increased, with a concentration - dependent and time-dependent manner,as compared with those in control group (P < 0. 05 ). Meanwhile,the expression levels of Bax,caspase-3 mRNA and protein were obviously up-regulated,but the expression levels of Bcl-2,survivin mRNA and protein were significantly down-regulated,with a concentration-dependent and time-dependent manner,as compared with those in control group (P < 0. 05). Conclusion The PBMNCs induced by IL-23 can inhibit cell proliferation of MUTZ-1 cells and can promote apoptosis of MUTZ-1 cell,and its action mechanism may be correlated with up-regulating the expressions of Bax and caspase-3 and down-regulating the expressions of Bcl-2 and survivin.%目的 白介素-23(IL-23)诱导人外周血单个核细胞对骨髓增生异常综合征(MDS)细胞系MUTZ-1增殖、凋亡及相关基因表达的影响.方法 采用密度梯度离心法分离获得正常人外周血单个核细胞(PBMNC),细胞分4组:阴性对照组(未加IL-23),3个浓度IL-23组(2、10、50 ng/ml),体外诱导72 h,并与MUTZ-1共培养.MTT实验检测MUTZ-1细胞增殖.流式细胞仪检测MUTZ-1细胞凋亡和细胞周期变化.Real time-PCR和Western-blot检测MUTZ-1细胞Bax、Bcl-2、caspase-3、survivin mRNA和蛋白表达.结果 2、10、50 ng/ml IL-23诱导的PBMNC与MUTZ-1细胞共培养后,MUTZ-1细胞增殖速度、S期细胞比例明显降低,细胞凋亡率、G0/G1期细胞比例明显增加,呈时间和浓度依赖性,与阴性对照组比较差异均有统计学意义(P<0.05);同时Bax、caspase-3 mRNA和蛋白表达明显上调,Bcl-2、survivin mRNA和蛋白表达明显下调,呈时间和浓度依赖性,与阴性对照组比较差异均有统计学意义(P<0.05).结论 IL-23诱导PBMNC能够明显抑制MUTZ-1细胞增殖,促进MUTZ-1细胞凋亡,其机制与上调Bax、caspase-3表达、下调Bcl-2、survivin表达有关.