Effect of Notch1 gene expression on the proliferation and apoptosis of T-cell acute lymphoblastic leukemia SupT1 cells and the mechanism
10.3760/cma.j.issn.1009-9921.2015.11.002
- VernacularTitle: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
- Author:
Qi YANG
;
Jianmin KANG
;
Xiuhua CHEN
;
Can LI
;
Fanggang REN
;
Yaofang ZHANG
;
Lihong HOU
- Publication Type:Journal Article
- Keywords:
Leukemia,lymphoblastic,acute;
Notch 1;
Apoptosis;
RNA interference
- From:
Journal of Leukemia & Lymphoma
2015;24(11):645-649
- CountryChina
- Language:Chinese
-
Abstract:
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 Notch1-shRNA gene and nonspecific Notch1-shRNA gene.The inhibitive rate of SupT1 cells was detected by CCK-8.The rates of early apoptotic cells (Annexin V+/ 7-AAD-) and late apoptotic cells (Annexin V+/7-AAD+) were analyzed by flow cytometry and the expression levels of Notch1 receptor gene and downstream target genes were assessed by quantitative reverse transcription and polymerase chain reaction (QT-PCR).Results The cell inhibition rates of Notch1 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 Notch1 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 Notch1 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 Notch1 receptor gene and its target genes (Hesl,c-myc,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 Notch1-shRNA can effectively decrease the Notch1 mRNA expression,and reduce the expression level of downstream target genes.Notch1 cut can inhibit the proliferation of SupT1 cells,and promote the early apoptosis.
