Fbxw7在急性T淋巴细胞白血病中的表达及其对细胞生长的抑制作用  

Expression of Fbxw7 in acute T lymphoblastic leukemia and its inhibitory effect on cell growth

作　　者：黄文强[1] 胡松元 靳松[3] 龙思方[1] 邹路红 严匡华 Huang Wenqiang;Hu Songyuan;Jin Song;Long Sifang;Zou Luhong;Yan Kuanghua(Department of Medical Laboratory,Qiannan Medical College for Nationalities;Department of Clinical Laboratory,The People’s Hospital of Qiannan/The First Affiliated Hospital of Qiannan Medical College for Nationalities;Department of Clinical Laboratory,Qiannan State Hospital of Traditional Chinese Medicine/The Second Affiliated Hospital of Qiannan Medical College for Nationalities;Department of Clinical Laboratory,The Third Affiliated Hospital of Guizhou Medical University;Department of Hematology,The First Affiliated Hospital of Henan University)

机构地区：[1]黔南民族医学高等专科学校医学检验系,都匀550639 [2]黔南州人民医院黔南民族医学高等专科学校第一附属医院检验科,都匀558000 [3]黔南州中医医院黔南民族医学高等专科学校第二附属医院检验科,都勻558000 [4]贵州医科大学第三附属医院检验科,都匀558000 [5]河南大学第一附属医院血液科,开封475001

出　　处：《重庆医科大学学报》2020年第2期257-262,共6页Journal of Chongqing Medical University

摘　　要：目的:观察F框/WD40域蛋白7(Fbxw7)在急性T淋巴细胞白血病中的表达,探讨其对细胞生长的影响及其机制。方法:采用逆转录聚合酶链反应(RT-PCR)检测62例急性T淋巴细胞白血病患者中的Fbxw7 mRNA表达情况,将携带Fbxw7的过表达慢病毒载体转至白血病Jurkat细胞后,采用RT-PCR和Western blot检测其转染效果,MTT法检测Jurkat细胞增殖情况,流式细胞仪检测细胞周期变化和细胞凋亡情况,Western blot检测c-Myc蛋白和周期蛋白E(Cyclin E)的表达。结果:急性T淋巴细胞白血病复发组患者和初发组患者中Fbxw7 mRNA表达水平较正常健康人明显降低(P<0.05);病毒转染后,Fbxw7组(转染携带Fbxw7过表达慢病毒载体)中Fbxw7 mRNA和蛋白质的相对表达水平较对照组(未转染的细胞)、阴性组(转染对照慢病毒载体)明显升高(P<0.05);转染24 h后,细胞增殖受影响不明显(P>0.05),而转染48 h和72 h后,细胞增殖明显受到抑制(P<0.05);转染48 h后,与对照组相比,Fbxw7组Jurka细胞中G0/G1期细胞所占比例和细胞凋亡率明显升高,S期和G2/M期细胞所占百分比、c-Myc和Cyclin E蛋白的相对表达量明显降低,差异均有统计学意义(P<0.05)。结论:Fbxw7在急性T淋巴细胞白血病中低表达,上调其表达可抑制白血病Jurkat细胞增殖,促进细胞凋亡,其作用机制可能与下调c-Myc和Cyclin E蛋白表达有关。Objective:To investigate the expression of F-box and WD40 domain-containing7(Fbxw7) in acute T lymphocyte leukemia and its effect on cell growth as well as its mechanism. Methods:The mRNA expression of Fbxw7 in 62 patients with acute T lymphocyte leukemia was detected by RT-PCR. The lentivirus vector carrying Fbxw7 overexpression was transferred to leukemia Jurkat cells and the effect of transfection was tested by RT-PCR and Western blot. And the proliferation of Jurkat cells was measured by MTT assay. The cell cycle and apoptosis were examined by flow cytometry,and the expression of c-Myc and Cyclin E protein was checked by Western blot. Results:Expression levels of Fbxw7 mRNA in relapse group and initial group were significantly lower than those in normal healthy group(P<0.05). After transfection,relative expression levels of Fbxw7 mRNA and protein in the Fbxw7 overexpression group(transfected with Fbxw7 overexpressed lentivirus vector)were significantly higher than those in the control group(no transfected cells) and negative control group(transfected with control lentivirus vector)(P<0.05). Cell proliferation was not significantly influenced(P>0.05) after transfection of 24 h,while was significantly inhibited after transfection of 48 h and 72 h(P<0.05). After transfection of 48 h,when compared with control and negative control groups,cell apoptosis and the proportion of cells in G0/G1 phase of Jurka cells were significantly increased in Fbxw 7 group,S phase and G2/M phase were significantly decreased,and relative expression levels of c-Myc and Cyclin E were significantly decreased,all with statistically significant difference(P<0.05).Conclusion:Fbxw7 is low expression in acute T lymphoblastic leukemia,its overexpression can inhibit the proliferation of Jurkat leukemia cells and promote cell apoptosis,and its mechanism may be related to the down-regulation of c-Myc and Cyclin E protein expression.

关 键 词：急性T淋巴细胞白血病 Fbxw7 细胞生长 抑制作用 

分 类 号：R733.71[医药卫生—肿瘤]