摘要
目的 检测外源性DLK1蛋白作用后急性T淋巴细胞白血病（T-ALL）细胞株CCRF-CEM（简称CEM）细胞的增殖情况及Notch信号通路下游靶基因的表达水平,探讨DLK1对CEM细胞中Notch信号通路的作用.方法 采用CCK-8法检测不同浓度（0.5、1.0、1.5μg/ml）DLK1蛋白作用后,不同时间点（24、48、72 h）CEM细胞的增殖情况；采用实时荧光定量-PCR法检测DLK1蛋白作用后不同时间点（24、48、72 h）CEM细胞中Notch信号通路的Notch1受体及其下游靶基因的mRNA表达水平.结果 DLK1蛋白促进CEM细胞增殖,与对照组相比差异具有统计学意义（P＜0.05）,其中不同蛋白浓度在72 h的增殖率分别为0.14±0.03、0.17±0.04、0.55±0.01.DLK1蛋白引起Notch信号通路的Notch1受体及其下游靶基因HES1、c-myc、NF-κB表达上调,与对照组相比差异具有统计学意义,其中靶基因HES1在72 h、c-myc在48 h、NF-κB在72 h的相对表达量分别为3.2551 ±0.3100、1.6086±0.0941、2.0515±0.3453.结论 外源性DLK1通过上调Notch信号通路的Notch1受体及其下游靶基因HES1、c-myc、NF-κB的表达,促进T-ALL细胞株CEM细胞增殖.
Objective To detect the proliferation and the expression levels of downstream target genes of Notch pathway of T-ALL CCRF-CEM（CEM）cell line treated with exogenous DLK1 protein,in order to investigatethe effects of DLK1 protein on the Notch pathway in CEM cells.Methods CCK-8 assay was performed to examine the proliferation of CCRF-CEM cells,which were treated with various concentration（0.5,1.0,1.5 μg/ml）DLK1 for various time（24,48,72 h）.RFQ-PCR was applied to assess the mRNA expression level of Notch1 receptor and downstream target genes of Notch pathway in CEM cells,which were treated for various time（24,48,72 h）.Results DLK1 protein stimulated the proliferation of CCRF-CEM cells,and the proliferation rates with different concentrations of DLK1 were 0.14±0.03,0.17±0.04,0.55±0.01 in 72 hours,respectively,there were statistic differences between that in the experimental group and that in the control group（P＜0.05）.DLK1 protein up-regulated the Notch1 receptor and its downstream target genes HES1,c-myc and NF-κB.The relative transcript levels of target genes HES1 in 72 hours,c-myc in 48 hours and NF-κB in 72 hours were 3.2551±0.3100,1.6086±0.0941,2.0515±0.3453 respectively,and there were statistic differences between that in the experimental group and that in the control group（P＜0.05）.Conclusion DLK1 protein stimulates the proliferation of T-ALL CCRF-CEM cells by up-regulating Notch1 receptor and downstream target genes HES1,c-myc and NF-κB of Notch pathway.
出处
《白血病．淋巴瘤》
CAS
2013年第10期586-588,596,共4页
Journal of Leukemia & Lymphoma
基金
山西省科技攻关项目（20120313020-7）
