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[Influence of Nucleostemin Expression Down-Regulation on PI3K/AKT/mTOR Signal Pathway in HL-60 Cells]
Yu Jia1, Yuan-Yu Wei2, Fan Zhang3
1Renal Laboratory of The First Afiliated Hospital of Zhengzhou University; Department of Laboratorial Medicine, The First Affiliated Hopital of Zhengzhou University, Henan Provincial Key Laboratory of Clinical Laboratory Medicine,Faculty of Laboratorial Medicine, Zhengzhou University; Henan Provincial Key Subject of Clinical Medicine, Zhengzhou 450052, Henan Province, China.
Objective:
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 leukemia HL-60 cells.
Methods:
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.
Results:
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(t1=2.31,P>0.05;t2=3.62,P>0.05;t3=1.60,P>0.05;t4=2.72,P>0.05) in comparison with control; the expression of GβL protein was statistically down-regnlated (t=15.01,P=0.002).
Conclusion:
The changes of GβL protein correlats with NS knockdown. The PI3K/AKT/mTOR pathway may be one of nucleostemin p53-independent signal pathways.
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