Effects Of 12-Epi-napeline On The Proliferation Of Leukemia Cells And Its Mechanisms

Objective:To investigate the inhibitory effect of 12-epi-napelline on K-562 and HL-60 cells of leukemia cells and its mechanism.Methods:The inhibitory effect of 12-epi-napelline on K-562 and HL-60 cells were evaluated by CCK-8 assay.The apoptosis and cell cycle effects of 12-epi-napelline on K-562 and HL-60 cells was detected by flow cytometry.The apoptosis morphology effect of 12-epi-napelline on K-562 and HL-60 cells was detected by Hoechst 33258.The effect of 12-epi-napelline on caspase-3,Bcl-2,PI3K,AKT,p-AKT and mTOR protein levels in K-562 and HL-60 cells were measured by Western blot.The effect of IGF-1 on apoptosis of K-562 and IIL-60 cells induced by 12-epi-napelline was detected by flow cytometry.The effect of IGF-1 on the inhibition of PI3K,AKT,P-AKT and mTOR protein expression by 12-epi-napelline in K-562 and HL-60 cells was detected by Western blot.Result:The CCK-8 method showed that 12-epi-napelline inhibited the proliferation of K-562 and HL-60 cells,and the survival rate decreased with the increase of the 12-epi-napelline concentration,compared with the control group,the difference was statistically significant(P<0.05),which was time-dose-dependent.The cell cycle effects of 12-epi-napelline on K-562 and HL-60 cells was detected by flow cytometry,the results showed that compared with the control group,12-epi-napelline group increased the percentage of G1 cells in K-562 and HL-60 cells,the difference was statistically significant(P<0.05).The apoptosis effects of 12-epi-napelline on K-562 and HL-60 cells was detected by flow cytometry and Hoechst staining,the results showed that compared with the control group,the apoptosis rate of K-562 and HL-60 cells in 12-epi-napelline group increased,and the apoptosis rate increased gradually with the increase of concentration,and the difference was statistically significant(P<0.05);And Western blot was used to detect apoptosis related proteins of K-562 and HL-60 cells,the results showed that the expression of Caspase-3,Bcl-2 was decreased by 12-epi-napelline.Furthermore,we explore its mechanism by Western blot,the results showed that 12-epi-napelline reduced the expression of PI3K,AKT,P-AKT and mTOR in K-562 and HL-60 cells.In addition,flow cytometry analysis showed that showed that IGF-1 inhibited the apoptosis induced by 12-epi-napelline in K-562 and HL-60 cells;Western blot analysis showed that IGF-1 inhibited the inhibitory effect of 12-epi-napelline on the expression of PI3K,AKT,P-AKT and mTOR in K-562 and HL-60 cells.Conclusion:12-epi-napelline can inhibit the proliferation,induce apoptosis and arrest the cell cycle of leukemia cells,and its mechanism may be related to the inhibition of PI3K/AKT/mTOR pathway.