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

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 mRNA expression levels of Notch1 receptor gene and its target genes (Hes1, 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. Key words: Leukemia, lymphoblastic, acute; Notch1; Apoptosis; RNA interference