CIBICI   14215
CENTRO DE INVESTIGACION EN BIOQUIMICA CLINICA E INMUNOLOGIA
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
B16 murine melanoma cells stably expressing shRNAMyD88
Autor/es:
NÚÑEZ, NICOLÁS; NOCERA, DAVID; RIVERO, VIRGINIA; MACCIONI,
Reunión:
Congreso; FIRST FRENCH-ARGENTINE IMMUNOLOGY CONGRESS. FAIC 2010; 2010
Institución organizadora:
Sociedad Argentina Inmunologia
Resumen:
Toll-like receptors (TLRs) recognize molecules derived from
pathogens (PAMPs) as well as endogenous danger signals possessing
similar chemical structures (DAMPs). Upon recognition
of their ligands, TLRs transduce signals through two pathways
involving distinct adaptors, Toll/IL-1R domaincontaining adaptor
inducing IFNs (TRIF) and Myeloid differentiation primary response
protein (MyD88), which is used by all TLRs except TLR3.
Stimulation of TLRs-pathways on antigen presenting cells has
always been considered a valuable tool to activate an anticancer
immune response. Recently, the presence of functional TLRs in
different tumor cell lines and in ex-vivo tumor samples has been
demonstrated, raising the question about the role that these
TLRs could be playing in sterile environments like tumors. Here,
we investigated whether MyD88-dependent signaling modulates
tumor development. The expression of MyD88 was stably
knocked down in the B16 melanoma cell line (B16-MyD88kd) by
using a silencing vector expressing short hairpin RNA (shRNA)
targeting mouse MyD88. As control, we generated B16 cells
stably transfected with the control vector encoding shRNA targeting
luciferase gene (B16-Luc). These vectors encode GFP fusion
protein for tracking the transfected cells. The expression of
MyD88 in B16-MyD88kd and in B16- Luc was evaluated by qRTPCR.
Approximately, a 65% of inhibition in mRNA expression
of MyD88 was observed in B16-MyD88kd. Interestingly, when
B16- MyD88kd were inoculated in vivo into syngeneic host,
the tumors elicited were bigger than those elicited by B16-Luc
(p<0.05). Our preliminary results indicate that in physiological
conditions, an endogenous ligand could ordinarily activate this
pathway and promoting a reduced tumor growth.