Effect of PKF118-310 on apoptosis of leukemia K562 cells and its mechanism

Abstract

Objective To investigate the effect of small molecule antagonists PKF118-310 on apoptosis of human chronic myeloid leukemia cell line K562 and its mechanism. Methods After the treatment of PKF118-310 at different concentration, the proliferation inhibition in K562 cells was detected by MTT, the β-catenin/TCF transcription complex in the nucleus was observed by immunofluorescence, the apoptosis was detected by flow cytometry, and the expressions of caspase-3, caspase-8, XIAP, bcl-2, β-catenin, TCF, c-myc and cyclin D1 were detected by Western blot. Results PKF118-310 inhibited the proliferation of K562 cells. The median inhibitory concentration (IC50) of PKF118-310-treated K562 cells for 24 h, 48 h and 72 h were 5.388, 3.290, 1.566 μmol/L, respectively. The β-catenin/TCF transcription complex in the nucleus was observed. After the treatment with 1.6 and 3.2 μmol/L PKF118-310 for 48 h, the apoptosis rates of K562 cells were (48.0±0.9) % and (80.2±1.2) %, respectively, and were higher than that in the control group [(1.2±0.6) %] (both P < 0.05). After the treatment of K562 cells with PKF118-310 at different concentration for 72 h, the expression levels of caspase-3 and caspase-8 were increased (both P < 0.05), and the expression levels of XIAP, bcl-2, β-catenin, TCF, c-myc and cyclin D1 were significantly decreased compared with control group (all P< 0.05). Conclusion PKF118-310 can inhibit K562 cells'proliferation, and induce the apoptosis. Key words: Leukemia, myeloid, chronic; K562 cells; Wnt/β-catenin signaling pathway; PKF118-310; Apoptosis