Effect of Sphk1-Specific Inhibitor PF543 on Apoptosis and Proliferation in Non-small Cell Lung Cancer Cells
Objective To investigate the effect of sphingosine kinase 1(Sphk1)inhibitor PF543 on apoptosis and proliferation of non-small cell lung cancer A549 cells.Methods Non-small cell lung cancer cell line A549 was cultured in vitro and collected.Different concentrations of PF543 were added to the collected A549 cells and recorded as group A(0 μmol·L-1),group B(10 μmol·L-1),group C(20 μmol·L-1),and then healthy human lung cells were selected as the control group.Reverse transcription polymerase chain reaction(RT-PCR)was used to detect the expression level of Sphk1 in each group.The proliferation ability of A549 cells under different concentrations of PF543 was detected by CCK8 kit.The apoptosis rate of A549 cells was detected by flow cytometry under different concentrations of PF543.Western blot was used to detect the relative expression levels of Caspase-3,Ki67,and Sphk1 in A549 cells under different concentrations of PF543.Results The expression level of Sphk1 mRNA in group A was higher than that in the control group(P＜0.05).The expression levels of Sphk1 mRNA in group B and group C were lower than those in group A,and group C was lower than group B(P＜0.05).At the same culture time,compared with group A,the tumor cell growth inhibition rates in both group B and group C increased,and group C was higher than group B(P＜0.05).At the same concentration,the longer the culture time,the higher the inhibition rate of tumor cell growth(P＜0.05).Compared with group A,the apoptosis rate of A549 cells in both group B and group C increased,and group C was lower than group B(P＜0.05).Compared with group A,the relative expression level of Caspase-3 in group B and group C increased,and group C was higher than group B(P＜0.05).The relative expression levels of Ki67 and Sphk1 in group B and group C decreased,and group C was lower than group B(P＜0.05).Conclusion Sphk1 is highly expressed in non-small cell lung cancer.PF543 can downregulate the expression level of Sphk1 in A549 cells,inhibit A549 cell proliferation,and promote A549 cell apoptosis.The mechanism of action may be that PF543 can upregulate the expression level of Caspase-3 to promote cell apoptosis,and downregulate the expression level of Ki67 to inhibit cell proliferation.
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