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Blood, Vol. 95 No. 2 (January 15), 2000:
pp. 510-518
Protein kinase C- isoform is involved in erythropoietin-induced
erythroid differentiation of CD34+ progenitor cells from
human bone marrow
June Helen Myklebust,
Erlend B. Smeland,
Dag Josefsen, and
Mouldy Sioud
From the Department of Immunology, The Norwegian Radium Hospital,
Oslo, Norway.
Protein kinase C (PKC) is a family of serine/threonine protein
kinases involved in many cellular responses. Although the analysis of
PKC activity in many systems has provided crucial insights to its
biologic function, the precise role of different isoforms on the
differentiation of normal hematopoietic progenitor cells into the
various lineages remains to be investigated. The authors have assessed
the state of activation and protein expression of PKC isoforms after
cytokine stimulation of CD34+ progenitor cells from human
bone marrow. Freshly isolated CD34+ cells were found to
express PKC- , PKC- 2, and PKC- , whereas PKC- , PKC- , and
PKC- were not detected. Treatment with erythropoietin (EPO) or with
EPO and stem cell factor (SCF) induced a predominantly erythroid
differentiation of CD34+ cells that was accompanied by
the up-regulation of PKC- and PKC-2 protein levels (11.8- and
2.5-fold, respectively) compared with cells cultured in medium.
Stimulation with EPO also resulted in the nuclear translocation of
PKC- and PKC-2 isoforms. Notably, none of the PKC isoforms tested
were detectable in CD34+ cells induced to myeloid
differentiation by G-CSF and SCF stimulation. The PKC inhibitors
staurosporine and calphostin C prevented EPO-induced erythroid
differentiation. Down-regulation of the PKC-, PKC-2, and PKC-
expression by TPA pretreatment, or the down-regulation of PKC- with
a specific ribozyme, also inhibited the EPO-induced erythroid
differentiation of CD34+ cells. No effect was seen with
PKC-2-specific ribozymes. Taken together, these findings point to a
novel role for the PKC- isoform in mediating EPO-induced erythroid
differentiation of the CD34+ progenitor cells from human
bone marrow.
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