Inhibition of crosstalk between Bcr-Abl and PKC signaling by PEITC, augments imatinib sensitivity in chronic myelogenous leukemia cells. (5th December 2015) - Roy, Madhumita
Abstract: Chronic myelogenous leukemia (CML), a clonal hyperproliferation of immature blood cells accounts for 20% of adult leukemia cases. Reciprocal translocation of chromosomes 9 and 22, results into Bcr-Abl fusion and is responsible for expression of a tyrosine kinase protein p210 bcr/abl, which mediates several survival pathways and confer therapeutic resistance. Protein kinase C (PKC), a family of serine threonine kinases play an important role in the process of leukemogenesis. A crosstalk between Bcr-Abl and PKC signaling has been documented. Therefore, targeting p210 bcr/abl and its associated signaling proteins using non-toxic natural means will be an effective strategy for antileukemic therapy. Aim of the present study is to investigate whether PEITC, a natural isothiocyanate in combination with imatinib mesylate (IM), a tyrosine kinase inhibitor could increase the therapeutic efficacy of IM by modulating the expression of p210 bcr/abl . Enhanced cytotoxic efficacy of IM by PEITC was further validated using another myelogenous leukemia cell line, KU812. It was observed that PEITC in combination with IM efficiently downregulated the expression of p210 bcr/abl in chronic myelogenous leukemia cell lines (K-562). PEITC inhibited the expressions of PKCα, PKCβII and PKCζ (both phosphorylated and total form). Expression of Raf1 and ERK1/2, two important target proteins in PKC signaling cascade was diminished. The result indicated that PEITC ultimately reduced expression of Raf1 and ERK1/2 through Bcr-Abl and PKC inhibition. This result was further confirmed by UCN-01, a selective PKC inhibitor and IM; indicating an association between p210 bcr/abl and PKC with Raf1 and ERK1/2. PEITC thus may have enormous potential in synergistic therapy of leukemia by enhancing drug efficacy. Highlights: PEITC increased efficacy of IM in K-562 as well KU812 cells. However, K-562 cells were more sensitive to IM in presence of PEITC. The extent of inhibition of p210 bcr/abl protein by imatinib mesylate was much more pronounced when cells were pretreated with PEITC. Expressions of total and phosphorylated PKC isoforms PKC (α, βII and ζ) were reduced by PEITC. PEITC inhibited the crosstalk between Bcr-Abl and PKC signaling as evident from the diminished expressions of Raf1and ERK1/2.