Notch1基因表达对急性T淋巴细胞白血病SupT1细胞增殖、凋亡的影响及其机制  

Effect of Notch1 gene expression on the proliferation and apoptosis of T-cell acute lymphoblastic leukemia SupT1 cells and the mechanism

作　　者：杨琦[1] 康建民[2] 陈秀花[3] 李参[2] 任方刚[3] 张耀芳[3] 侯丽虹[2] 

机构地区：[1]山西医科大学研究生学院,太原030001 [2]山西医科大学第二医院血液科 [3]山西医科大学第二医院血液科实验室

出　　处：《白血病．淋巴瘤》2015年第11期645-649,共5页Journal of Leukemia & Lymphoma

基　　金：山西省科技攻关项目（20120313020-7）

摘　　要：目的探讨Notch信号通路与急性T淋巴细胞白血病发生、发展的关系。方法利用携带Notch1特异性和非特异性shRNA的慢病毒载体包装成的病毒颗粒感染急性T淋巴细胞白血病SupT1细胞，采用CCK．8法检测细胞抑制率；采用流式细胞术检测细胞凋亡率（AnnexinV＋／7-AAD一和AnnexinV＋／7-AAD＋）；采用实时荧光定量PCR法检测Notch1受体基因及下游靶基因的表达水平。结果Notch1干扰组、空白对照组和空载体组96h的细胞增殖抑制率分别为0．902±0．013、0、0．486±0．084，干扰组较空白对照组、空载体组升高（均P〈0．05）；三组细胞早期凋亡率分别为（15．27±0．31）％、（5．57±0．25）％、（5．80±0．20）％，干扰组较空白对照组、空载体组升高（均P〈0．05）；而各组细胞晚期凋亡率无明显变化（均P〉0．05）。干扰组细胞中Notch1受体基因及其下游靶基因Hes1、c-myc、NF-κB在48、72、96h的mRNA相对表达量均较空白对照组及空载体组降低（均P〈0．05）。结论特异性Notch1-shRNA可有效下调Notch1mRNA的表达，并降低其下游靶基因的表达水平。Notch1表达下调可以抑制急性T淋巴细胞白血病SupT1细胞增殖，促进supT1细胞的早期凋亡。Objective To explore the relationship between T-cell acute lymphoblastic leukemia and the Notch signaling pathway. Methods Human T-cell acute lymphoblastic leukemia SupT1 cells were infected with the lentiviral vector made up specific Notchl-shRNA gene and nonspecific Notchl-shRNA gene. The inhibitive rate of SupT1 cells was detected by CCK-8. The rates of early apoptotie cells （Annexin V＋/ 7-AAD-） and late apoptotic cells （Annexin V＋/7-AAD＋） were analyzed by flow cytometry and the expression levels of Notchl receptor gene and downstream target genes were assessed by quantitative reverse transcription and polymerase chain reaction （QT-PCR）. Results The cell inhibition rates of Notchl interference group, control group and empty vector group at 96 h were 0.902±0.013, 0, and 0.486±0.084, respectively, and it was increased obviously in Notehl interference group （both P 〈 0.05）. The cell early apoptosis rates of the three groups were （15.27±0.31） %, （5.57±0.25） %, （5.80±0.20） %, respectively. The cell early apoptosis rate of Notehl interference group was increased obviously compared with the control group and empty vector group （both P 〈 0.05）. While the cell late apoptosis rates had no significant difference among the three groups （P 〉 0.05）. The mRNA expression levels of Notchl receptor gene and its target genes （Hesl, c-mye, NF-κB） at 48 h, 72 h and 96 h were higher than those in the control group and empty vector group （all P 〈 0.05）. Conclusions The specificity of Notehl-shRNA can effectively decrease the Notchl mRNA expression, and reduce the expression level of downstream target genes. Notchl cut can inhibit the proliferation of SupT1 cells, and promote the early apoptosis.

关 键 词：白血病 淋巴细胞 急性 NOTCH1 细胞凋亡 RNA干扰 

分 类 号：R733.71[医药卫生—肿瘤]