Effect Of Notch1 Expression On The Proliferation And Bortezomib Sensitivity Of T- ALL Cells

Objective:To detect Notch1 genes by interference after Sup T1 cell proliferation, apoptosis,Notch1 receptor gene and its downstream target genes on the expression level of change,explore the development and the relationship of the Notch signaling pathway and acute T lymphocyte leukemia.After sh RNA role of Sup T1 cells to add 1 ng/μl with meters for boron, detection of Sup T1 cells proliferation, apoptosis, and the expression of the gene of Notch1 recerptor and downstream target genes of Notch1 recerptor, and explored the effect of Notch1 gene expression of bortezomib sensitivity.Methods:1.Human Sup T1 cells were infected with specific Notch1-sh RNA and nonspecific Notch1-sh RNA Lentiviral Vector, selecting interference efficiency of infected cells as interference group. CCK-8 was used to detect the proliferation of Sup T1 cells;The percentages of early apoptotic cells(Annexin V+/7-AAD-)and late apoptotic cells(Annexin V+/7-AAD+)were analyzed by flow cytometry;Quantitative reverse transcrip-tion and polymerase Chain reaction(QT-PCR) was applied to assess the Notch1 receptor and downstream target genes Hes1, NF-κB, c-myc m RNA expression level at 48, 72, 96 hours, respectively.2.After jioning 1 ng/μl bortezomib in infected at 24, 48, 72 hours, respectively and incubating the Sup T1 cells for 24 hours, we detected proliferation, apoptosis and Notch1 receptor gene and downstream target genes Hes1, NF-κB, c-myc m RNA expression level of the Sup T1 cells by the CCK-8 method, flow cytometry, real-time fluorescentquantitative PCR method at 48, 72, 96 hours, respectively.Results:1.Proliferation of Notch1 interference group cell significantly reduced comparing with blank group and the empty carrier group, the inhibition rate of interference group increased significantly comparing with blank group and the empty carrier group(P<0.05),and the inhibition showed time-dependence.2.The Annexin V-PE+/7-AAD- of interference group compared with blank control group and the empty carrier group increased significantly(P<0.05); and the Annexin V-PE+/7-AAD+of each group was no significant change(P>0.05).3.Notch1 receptor gene of interference group and downstream target genes of Notch1 recerptor Hes1, c-myc, NF-κB compared with blank group and the empty expression vector group decreased significantly(P<0.05).4.Bortezomib inhibited the proliferation of Sup T1 cells obviously, and inhibition showed concentration and time dependence;the inhibition rate of interference and Bortezomib group and empty vector and Bortezomib group were higher than interference group and empty vector group(P<0.05).5.The Annexin V-PE+/7-AAD- of interference and Bortezomib group comparing with blank group and empty carrier group increased significantly(P<0.05); and the Annexin V-PE+/7-AAD+of each group was no significant change(P>0.05).6.Notch1 receptor gene of interference and Bortezomib group and downstream target genes of Notch1 recerptor Hes1, c-myc, NF-κB comparing with interference group significantly decreased(P<0.05), and the inhibition showed time-dependence.Conclusions:1.Specificity Notch1- sh RNA- 4252 can effectively down regulated the Notch1 m RNA expression, and reduce the expression of downstream target genes.2.Notchl down regulated can inhibit the Sup T1 cells proliferation, promote earlyapoptosis Sup T1 cells, and the inhibition showed time-dependence.3.The inhibition of Sup T1 cells infected by sh RNA and affected by bortezomib proliferation and apoptosis,Notch1 receptor gene and the downstream target genes was more notable, Sh RNA and Bortezomib showed overlaping function.