Differentiation of myeloid cell lines correlates with a selective expression of RIZ protein.
BACKGROUND
The retinoblastoma-interacting zinc-finger gene RIZ is expressed in two forms (RIZ1 and RIZ2) that differ for the presence near the N-terminus of RIZ1 of a conserved domain, defined PR (PRDI-BF1-RIZ homology), homologous to a similar domain present in other proteins recognized as tumor suppressor gene products. The RIZ1 form is usually absent or expressed at low levels in tumor cells, whereas RIZ2 is frequently expressed. We investigated a possible involvement of RIZ1 in differentiation control using a myeloid cell maturation model that is easily modulated by retinoids and other agents.


METHODS
HL60 or NB4 cell lines or patients' leukemic promyelocytes were treated with all- trans -retinoic acid or other agents to induce differentiation. RIZ gene expression was determined with reverse transcriptase polymerase chain reaction (RT-PCR) and RNase protection assay. Immunocytochemistry was performed to assess variation of the intracellular distribution of RIZ protein on all- trans-retinoic acid treatment. Forced expression of RIZ1 protein was obtained with a recombinant adenovirus containing RIZ1 cDNA.


RESULTS
Treatment with retinoic acid induced a selective expression of RIZ1 in HL60 cell line. Retinoic acid effect was maximal at 7 days and correlated to the granulocytic differentiation of cells. A similar effect was obtained in retinoic acid-sensitive NB4 cell line or in patients' leukemic promyelocytes, but not in the retinoic acid-resistant cell line NB4.007/6 or in the U937 cell line. Selective expression of RIZ1 was also induced by 12-O-tetradecanoyl-phorbol-13-acetate in the U937 and HL60 cell lines and by 1,25-dihydroxyvitamin D(3) only in HL60 cells. In HL60 cells, RIZ1 was also induced by activation of a retinoid alpha receptor-independent maturation pathway based on retinoid X receptor agonist and protein kinase A synergism. In addition, retinoic acid produced a redistribution of the antigen within the nucleus in these cells. Forced expression of RIZ1 protein induced growth arrest and death of HL60 cells.


CONCLUSIONS
The correlation between the selective expression of RIZ1 induced by retinoic acid, 12-O-tetradecanoyl-phorbol-13-acetate, or 1,25-dihydroxyvitamin D(3) and differentiation suggested that RIZ protein was involved in myeloid cell differentiation induced by these agents.
Gazzerro
P
P
Istituto di Patologia generale ed Oncologia, Seconda Università degli studi di Napoli, Naples, Italy.
Bontempo
P
P
Schiavone
E M
EM
Abbondanza
C
C
Moncharmont
B
B
Armetta
I
I
Medici
N
N
De Simone
M
M
Nola
E
E
Puca
G A
GA
Molinari
A M
AM
D000249:Adenosine Monophosphate / Q000031:analogs & derivatives; D000256:Adenoviridae / Q000378:metabolism; D000970:Antineoplastic Agents / Q000494:pharmacology; D001565:Benzoates / Q000494:pharmacology; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D004268:DNA-Binding Proteins; D011495:Histone-Lysine N-Methyltransferase; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D022423:Myeloid Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D016160:Retinoblastoma Protein / Q000235:genetics / Q000378:metabolism; D012176:Retinoids / Q000494:pharmacology; D014157:Transcription Factors; D014212:Tretinoin / Q000494:pharmacology; D016335:Zinc Fingers / Q000235:genetics
D000970:Antineoplastic Agents; D001565:Benzoates; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D016160:Retinoblastoma Protein; D012176:Retinoids; D014157:Transcription Factors; C087130:SR 11237; D000249:Adenosine Monophosphate; D014212:Tretinoin; D011495:Histone-Lysine N-Methyltransferase; C093563:PRDM2 protein, human