抑制mel18表达增强桂皮醛诱导的HL60细胞分化  

Suppression of mel18 enhances differentiation of acute myeloid leukemia cell line HL60 induced by cinnamaldehyde

作　　者：刘黎琼[1] 刘泽林[1] 崔海燕[1] 金梦迪[1] 

机构地区：[1]深圳市南山区人民医院血液科,广东深圳518052

出　　处：《中国病理生理杂志》2017年第10期1852-1857,共6页Chinese Journal of Pathophysiology

基　　金：广东省自然科学基金资助项目(No.2016A030313032);深圳市科技研发资金基础研究项目(No.JCYJ20160429182558793);深圳市科技局资助项目(No.201303188);深圳市南山区卫生计划项目(No.2015001)

摘　　要：目的:研究下调mel18基因表达对桂皮醛(CA)诱导HL60细胞分化的影响。方法:用低浓度CA和反式维甲酸(ATRA)处理HL60细胞。用shRNAmel18质粒及对照质粒shRNALuc包装慢病毒,用病毒感染HL60细胞。低浓度CA和ATRA作用于病毒感染的HL60细胞,流式细胞术检测细胞周期和细胞表面分化抗原表达,Western blot法检测MEL18蛋白、cyclin D1、p27的表达和Akt的磷酸化水平。结果:低浓度CA和ATRA增加HL60细胞的粒细胞分化抗原CD11b表达,MEL18蛋白表达下降。shmel18病毒感染的HL60细胞(shmel18-HL60细胞)MEL18蛋白表达下降,而对照病毒感染的HL60细胞(sh Luc-HL60细胞)MEL18蛋白表达无明显变化。shmel18-HL60细胞的CD11b表达率明显增高,细胞阻滞于G1期;经低浓度CA处理后,shmel18-HL60细胞的CD11b表达率进一步增高。shmel18-HL60细胞Akt的磷酸化水平及cyclin D1表达明显下降,而p27表达则显著升高。结论:抑制mel18基因表达导致HL60细胞向成熟粒细胞方向分化,mel18基因可能通过PI3K/Akt信号途径调控cyclin D1和p27的表达,影响HL60细胞分化。而CA可能通过抑制mel18基因表达从而通过PI3K/Akt途径促进HL60细胞分化。AIM：To investigate the effect of suppression of mel18 gene on the differentiation of human acute myeloid leukemia cell line HL 60 induced by cinnamaldehyde （ CA） .METHODS：HL60 cells were treated with low con-centration of CA or all-trans retinoic acid （ATRA）.shRNAmel18 vector and shRNALuc control vector were employed to package lentiviruses which were then used to infect HL 60 cells.The virus-infected HL60 cells were treated with low con-centration of CA , and ATRA was used as a positive control of differentiation-inducing agent .The differentiation markers on the cell surface and cell cycle of virus-infected HL60 cells were analyzed by flow cytometry .Western blot was used to deter-mine the expression of MEL18, cyclin D1 and p27, as well as the phosphorylation level of Akt .RESULTS：Low concen-tration of CA and ATRA increased the expression of granulocytic differentiation marker CD 11b on the HL60 cells, with the decreased expression of MEL 18 in the HL60 cells.The expression of MEL18 decreased in shmel18 virus-infected HL60 cells （shmel18-HL60 cells）, but did not change in shLuc-HL60 cells.The expression of CD11b on shmel18-HL60 cells increased with G 1-phase arrest , which went even higher after treatment with CA .The phosphorylation level of Akt and the expression of cyclin D1 decreased in shmel18-HL60 cells with the increase in the expression of p27.CONCLUSION：In-hibition of mel18 gene leads HL60 cell granulocytic differentiation .mel18 gene may affect the differentiation of HL 60 cells by regulating cyclin D1 and p27 via PI3K/Akt signaling pathway.PI3K/Akt signaling pathway is also involved in CA-in-duced differentiation of HL60 cells by suppressing mel18 gene expression.

关 键 词：桂皮醛 mel18基因 HL60细胞 细胞分化 PI3K/AKT信号通路 

分 类 号：R329.21[医药卫生—人体解剖和组织胚胎学] R733.7[医药卫生—基础医学]