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Effect of PI3K/Akt/mTOR signaling pathway inhibitors on proliferation and PHLPP protein expression of leukemia cell lines
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Abstract:
Objective: To observe the effect of PI3K/Akt/mTOR pathway inhibitors wortmannin or rapamycin on the proliferation and PHLPP (PH domain leucine-rich repeat protein phosphatase) protein expression of leukemia cell lines. Methods: Human leukemia cell lines K562 and HL-60 were treated with different concentrations of wortmannin or rapamycin. The proliferation of K562 and HL-60 cells was examined by WST-1 assay and the apoptosis of K562 and HL-60 cells was detected by Annexin Ⅴ-FITC double staining flow cytometry. The expressions of PHLPP, phosphorate-Akt (p-Akt) and total Akt protein were detected by Western blotting. Results: Wortmannin inhibited the proliferation of K562 and HL-60 cells in a dose- and time-dependent manner, with the IC50 value of 48 h being (187.6±48.4) nmol/L and (185.5±48.1) nmol/L (P<0.05). After treating K562 cells with 100 nmol/L wortmannin and HL-60 with 50 nmol/L wortmannin for 12 and 24 h, the apoptosis rates were significantly higher than those in the control group (\[12.4±0.7\]%, \[176±2.3\]% vs \[5.0±0.6\]%, P<0.05; \[11.0±0.2\]%, \[17.9±1.6\]% vs \[6.8±0.4\]%, P<0.05). After treating K562 and HL-60 cells with wortmannin for 12，24 and 36 h，the protein expressions of p-Akt and PHLPP were significantly reduced. And rapamycin also down-regulated the protein expression of PHLPP in K562 and HL-60 cells. Conclusion: The inhibitor of PI3K/Akt/mTOR signaling pathway inhibit the proliferation as well as PHLPP protein expression of leukemia cell lines.
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Project Supported:
Project supported by the Natural Science Foundation of Liaoning Province (No.20072105)
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