Effect of tetrabutyl propanediamine on human acute myeloid leukemia cell line HL-60 / 实用医学杂志

Objective To investigate the effect of the new polyamine analogue tetrabutyl propanediamine (TBP) on cell proliferation and the underlying mechanism. Methods MTT assay was performed to determine cell proliferation. Flow cytometry was performed to detect cell cycle transition. DNA fragmentation and mitochondrial membrane potential determinations were performed to detect cell apoptosis. The activity of key enzymes in polyamine catabolism was detected by chemiluminescence assay. Results TBP could significantly inhibit the proliferation of HL-60 cells by blockingcell cycle transition and by inducing apoptosis. The TBP-induced apoptosis of HL-60 cells was in a dose-dependent manner. The enzyme activities of SMO and APAO were also significantly increased in HL-60 cells after treatment with 100 μM TBP for 24 hours. Conclusions TBP, as a new putrescine analogue, could inhibit proliferation of HL-60 cells by increasing the enzyme activity of SMO and APAO and inducing apoptosis.

Inhibition of spermine oxidase expression by siRNA decreases sensitivity of human A549 lung cancer line to antitumor polyamine analogue CPENSpm / 中国药理学通报

Aim To evaluate the effect of expression inhibition of spermine oxidase(SMO)on the actitumor activity of polyamine analogue CPENSpm (N~1-cyclopropylmethyl-N~(11)-ethylnorspermine).Methods siRNA technique was used to inhibit expression of SMO in human lung cancer line A549.QT-RT-PCR and enzyme activity assay was performed to determine the expression level of SMO.The cell proliferation was detected by MTT assay.The apoptosis of A549 cells were evaluated by DNA degradation and Sub-G_1/flow cytometry assay.Results The A549 cell line with silenced SMO expression was successfully obtained.Basic SMO mRNA and enzyme activity levels in the SMO-siRNA plasmid transfected cells were 0.53% and 14% lower than that in the control cells respectively. Treating A549 control cells by 10 μmol·L~(-1) CPENSpm for 24 hours resulted in a 10-folds up-regulation of SMO in mRNA level and 20-fold increase in enzyme activity,but this drug-induced SMO expression was obviously prevented in SMO-siRNA plasmid transfected cells.MTT assay demonstrated that SMO expression inhibition decreased the sensitivity of A549 cells to CPENSpm exposure(0～20 μmol·L~(-1)).DNA degradation and sub-G_1 assay proved a deceased ability of CPENSpm to induce apoptosis in SMO-siRNA plasmid transfected cells.Conclusion Up-regulation of SMO by CPENSpm is possibly one of the molecular basics for its antitumor activity.

Polyamine analogue CPENSpm inhibits proliferation of human lung cancer cells by interfering polyamine metabolism / 中国药理学通报

Aim To study the effects of polyamine analogue CPENSpm on the human lung cancer line A549 in cell proliferation and apoptosis.Methods MTS was used to assay the cell proliferation,chemical analysis methods were used to determine the activities of enzymes in the polyamine metabolism,HPLC was performed to assay the intracellular concentration of polyamines,Sub-G1 and DNA fragmentation assays were used to determine the cell apoptosis.Results Treating A549 lung cancer cells by CPENSpm resulted in:①cell-growth inhibition and cell apoptosis;②inhibition of ODC(key enzyme in polyamine synthetic pathway)and activation of SSAT and SMO(key enzymes in polyamine catabolism);③great decrease of intracellular polyamine concentrations.MDL72527,the SMO inhibitor,can antagonize the effect of CPENSpm on inhibiting the proliferation of A549 cells.Conclusion CPENSpm inhibits proliferation and induces apoptosis of human A549 lung cancer cell line by interfering the polyamine metabolism,depleting intracellular polyamine contents that are need by quick-growth of cancer cells and inducing production of H2O2.