Carbenoxolone induces apoptosis and inhibits survivin and survivin-AEx3 genes expression in human leukemia K562 cells

ABSTRACT

Leukemia is a malignant disorder of the blood progenitor/stem cells which is characterized by abnormal proliferation of white blood cells. Although anti-cancer drugs induce apoptosis in cancerous cells, drug resistance is the significant problem mainly due to over-expression of inhibitors of apoptosis proteins [lAPs] such as survivin. In this content, it has been reported that an anti-inflammatory drug, Carbenoxolone [CBX], could induce apoptosis and growth inhibition in several types of cancerous cells. In the present study, effects of CBX on apoptosis and level of the expression of survivin gene and its deltaEx3 splicing variant have were evaluated in K562 cells. K562 cells were cultured and treated with different concentrations of CBX [50-300 microM] at different time intervals [12-48 hrs]. Trypan blue exclusion test was used to evaluate cell viability. Fluorescent microscopy [Acridine Orange/Ethidium Bromide double staining] and DNA fragmentation assay were used to study apoptosis. The expression level of survivin and its deltaEx3 splice variant were studied by RT- PCR. It was found that both growth inhibition and apoptosis occurred in K562 cells. In addition, down-regulation of survivin and survin-deltaEx3 were observed, after 2-4 hrs treatment with 150 microM of CBX. However, the expression level of survivin and its deltaEx3 splice variant increased in subsequent time [6-12 hrs] nearly to the level of control cells. From the results of this study, it may be concluded that CBX can be considered as a candidate for further studies in CML treatment, especially in the case of drug- resistant leukemia cells