Study The Effect On Chemosensitization Of Human Acute Monocytic Leukemic Cell Line SHI-1 Through SiRNA Mediated Ets2 Gene Silencing

Objective: To study the effect on chemosensitization of human acute monocytic leukemic cell line SHI-1 through siRNA mediated ets2 gene silencing and the Ets2 gene expression level in leukemia patients.Methods:â‘ . To detect expression levels of Ets2 gene in bone marrow of 28 patients with leukemia and SHI-1 cell line by RT-PCR, taking the normal blood cells of 5 the healthy as control.â‘¡To designe and chemical synthesise three short interfering RNA (siRNA) targeting Ets2 gene. Electroporation method was used for cell transfection of a Ets2 siRNA. Non-transfected cells and Non-specifical siRNA transfected cells were negative control groups. FAM-siRNA transfected cells were positive control. Transfection efficiencies were deceted by Flow Cytometry. Ets2 mRNA level was determined by Real-time PCR. Cell apoptosis ratio was measured by Annexin V-FITC/PI Flow Cytometry.Results:â‘ The results indicated that the high-expression of Ets2 gene in SHI-1 cell; Out of total 28 patients, Ets2 gene in bone marrow was successfully amplified from 21 cases ndd 14 in 19 (73.68%) ANLL patients and 3 in 5(60%) ALL patients expressed Ets2 gene.1 in 5(10%) the healthy expressed Ets2 gene. The expression ratio of healthy In addion, it wasn't also showed the correlation between the expression of Ets2 gene and sex; age; white blood cell count; hemoglobin level; platelet count or WHO classification of ANLL, possibly due to the small number of specimens.â‘¡The transfection efficiency was about 35%. The Ets2 siRNA-3 could significantly decrease the Ets2 gene mRNA level about 30%, while Ets2 siRNA-1 decreased mRNA level about 10%. We didn't find the inhibitory effect of Ets2 siRNA-2. siRNA mediated Ets2 gene silencing increased apoptosis rate to some extent(P=0.0001)and promoted VP-16-induced cell apoptosis(P=0.004).Conclusions: The high-expression of Ets2 transcription factor in leukemia cells was to promote growth and survival of leukemia cells. SiRNA mediated Ets2 gene silencing induced cell apoptosis and enhanced in vitro sensitivity to chemotherapy (VP-16) of Human Acute Monocytic Leukemic Cell Line SHI-1. It speculated the high-expression of Ets2 may actually be an unfavorable determinant of chemotherapy sensitivity in leukemia.