Erythropoietin Signaling in Tumor Cells: Differential Activation of Mitogen Activated Protein Kineses and Their Effect upon Cell Survival.

Erythropoietin (EPO) regulates the proliferation and differentiation of erythroid progenitors via its receptor, EPO-R and through various Mitogen activated protein kinase (MAPK) pathways. The role of EPO on other cell types is still unkown. In the present study we have elected to examine this aspect in a transformed pancreatic cell line, AR42J cells. We investigated the activation of two MAPKs, namely extracellular regulated kinase (ERK) and c-jun NH2-terminal kinase (JNK) after exposure of AR42J cells to 1 U/ml EPO using the Western blot analysis. We also examined cell viability during EPO exposure by high sensitivity fluoremetric method using CCK8. We found a rapid activation of ERK-1/2 in AR42J cells reaching the maximum of 3.3 fold in 5 min, while it took 30 min for JNK-1/2 to reach the maximum. In the absence of EPO, addition of specific JNK inhibitor, SP600125, reduced cell viability to 50% at a dose of 20 μM while with ERK inhibitor, UO126, cell viability was not reduced even up to 60 μM of the drug. To examine the effect of induction of MAPK by EPO on AR42J cell survival, cells were treated with inhibitors to ERK or JNK 1h prior to EPO addition and the cumulative cell survival were calculated from day 1 through 4. EPO addition to AR42J cells resulted in significantly higher cumulative cell survival of 1.00 ± 0.04 unit absorption compared to the value of 0.35 ± 0.02 unit absorption seen in controls without EPO. When cells were treated with EPO and ERK inhibitor a significantly higher cumulative cell survival of 1.50 ± 0.04 unit absorption was observed (p < 0.01) indicating ERK to be less effective in their survival. On the other hand, samples treated with EPO and JNK inhibitor had significantly lower cell survival (0.55 ± 0.06 unit absorption, p < 0.01) than the EPO control indicating an essential role of JNK in their survival. These results indicate that in EPO mediated survival of AR42J cells, activation of JNK appears to be more important than ERK activation thus indicating a key role of JNK in cell viability. It is very important to understand the role of EPO signaling in the viability of transformed cells since EPO is often supplemented after bone marrow transplantation to enhance the recovery after chemotherapy.