Sodium caseinate induces expression and secretion of murine multipotent myeloid cell line 32D macrophage colony-stimulating factor.
BACKGROUND
Evidence that sodium caseinate (CasNa) is capable of inhibiting proliferation of hematopoietic precursor cell line 32D and inducing its differentiation into macrophage cells has recently been published. Taking into consideration that hematopoiesis is regulated by growth factors and that macrophage colony-stimulating factor (M-CSF) is a well-known growth factor that induces differentiation of macrophages, in this work we evaluated whether CasNa is capable of inducing expression and secretion of M-CSF in 32D cells.


METHODS
We cultured 32D cells in presence and absence of CasNa and compared their proliferation and viability. RNA was extracted from cell lysates to evaluate expression of the gene for M-CSF and its receptor. Cultured conditioned media was used to evaluate presence of M-CSF.


RESULTS
Our results showed that CasNa inhibited proliferation of 32D cells and that conditioned media (CM) of these cultures contained M-CSF-like activity. Presence of M-CSF in CM was detected by inhibiting M-CSF activity with anti-M-CSF and presence of this growth factor was confirmed by ELISA assay. We also provided evidence that CasNa induced expression of mRNA for M-CSF in 32D cells as well as increased expression of mRNA for its receptor.


CONCLUSIONS
CasNa inhibits proliferation of 32D cells and induces expression of the gene for M-CSF and that of its receptor. It also induces secretion of the bioactive form of M-CSF.
Ramos
Gerardo
G
Laboratorio de Biología Celular y Molecular del Cáncer (L-324), Facultad de Estudios Superiores (FES)-Zaragoza, Universidad Nacional Autónoma de México, Calle Batalla 5 de Mayo s/n, Mexico City, 09230 Mexico.
Weiss
Benny
B
Córdova
Yolanda
Y
Hernández
Jorge
J
Zambrano
Isaac
I
Santiago
Edelmiro
E
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D002364:Caseins / Q000494:pharmacology*; D002454:Cell Differentiation; D002455:Cell Division; D002460:Cell Line; D045744:Cell Line, Tumor; D002614:Chelating Agents / Q000494:pharmacology; D003114:Colony-Forming Units Assay; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D006412:Hematopoietic Stem Cells / Q000166:cytology; D007377:Interleukin-3 / Q000378:metabolism; D016173:Macrophage Colony-Stimulating Factor / Q000378:metabolism*; D051379:Mice; D023461:Myeloid Progenitor Cells / Q000378:metabolism*; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D015854:Up-Regulation
D002364:Caseins; D002614:Chelating Agents; D017077:Culture Media, Conditioned; D007377:Interleukin-3; D012333:RNA, Messenger; D012313:RNA; D016173:Macrophage Colony-Stimulating Factor