Effects of Silencing AKT Gene by shRNA on Proliferation, Apoptosis and Notch1 Signal Pathway in Jeko-1 Cells

Abstract

To investigate the effects of silencing AKT gene by RNA interference on the proliferation, apoptosis and the expression of Notch1 signal pathway-related proteins in mantle cell lymphoma Jeko-1 cell line. The hairpin-like oligonucleotide sequences targeting AKT gene were designed and transfected into Jeko-1 cells by lipofectamine(TM) 2000. The AKT mRNA and protein were detected by RQ-PCR and Western blot respectively. Cell growth was determined by MTT. Cell apoptosis was analyzed by flow cytometry. The expression of BCL-2, BAX, caspase-3, caspase-9, p-AKT, Notch1, HES1 was detected by Western blot. AKT mRNA was markedly suppressed by the shRNA targeting AKT. AKT shRNA suppressed the proliferation of Jeko-1 cells and induced apoptosis of these cells. The cell apoptotic rates were (37.72±4.39)%, (2.62±1.53)%, (1.57±0.42)% in AKT shRNA, Neg-shRNA and Control, respectively, The difference between them was statistically significant (P<0.01). AKT shRNA down-regulated the expression of Bcl-2, AKT p-AKT , Notch1, and HES1, up-regulated the expression of BAX, caspase-3, caspase-9. Silencing AKT gene can inhibit the proliferation of Jeko-1 cells line, induc cell apoptosis, its mechanism may be associated with specially inhibiting PI3K/AKT signaling pathway and down-regulating the activity of Notch1 signaling pathway.