<p><b>OBJECTIVE</b>To explore the effect of nucleostemin(NS) RNAi on the expression of signal molecules in PI3K/AKT/mTOR pathway, a candidate of p53-independent signal pathway in the leukemiaHL-60 cells.</p><p><b>METHODS</b>The expression of NS was interfered by transfection of P53-deficient HL-60 cells with the recombinant lentivirus expression vector NS-RNAi-GV248. The exression of NS and signal molecules of PI3K/AKT/mTOR pathway were detected by Western blot.</p><p><b>RESULTS</b>The fluorescence microscopy showed that the recombinant lentivirus vector NS-RNAi-GV248 transfected HL-60 cells successfully with a 80% transfection rate. Western blot showed that the expression of NS protein was inhibited obviously in HL-60 cells, and the expression levels of AKT, p-AKT, p70s6k and p-p70s6k were not statistically different(t=2.31,P>0.05;t=3.62,P>0.05;t=1.60,P>0.05;t=2.72,P>0.05) in comparison with control; the expression of GβL protein was statistically down-regnlated (t=15.01,P=0.002).</p><p><b>CONCLUSION</b>The changes of GβL protein correlats with NS knockdown. The PI3K/AKT/mTOR pathway may be one of nucleostemin p53-independent signal pathways.</p>