Effects of oligodeoxynucleotide IMT504 on cell viability and gene expresion in a murine beta cell line (MIN6-B1)

CONVERTI A; MONTANER A; LIBERTUN C; LUX-LANTOS V; BONAVENTURA MM

Buenos Aires

Jornada; Reunión Conjunta de Sociedades de Biociencias; 2017

EFFECTS OF OLIGODEOXYNUCLEOTIDE IMT504ON CELL VIABILITY AND GENE EXPRESION IN A MURINE BETA CELL LINE (MIN6B1)Ayelén Converti (1), Alejandro Montaner, Carlos Libertun,Victoria Lux-Lantos, María Marta Bonaventura (1)(1) IBYME-CONICETWe have previously demonstrated that treatment with IMT504 promotes significant improvement in the diabetic condition in diverseanimal models. We have also shown effects on gene expressionon freshly isolated islets from diabetic IMT504-treated animals. Based on these results, here we evaluated if the effects of IMT504observed in vivo were due to direct effects on beta cell function. Inparticular we studied cell viability and gene expression. A murinebeta cell line (MIN6B1, kindly donated by Dr. P. Halban) was used.Cells were cultured in DMEM with 20 mM glucose, 15% SFB, 71 uMβmercaptoethanol. Cell viability was analized by MTS while geneexpression of Pdx-1, INS2, INS1 and MafA was analized by qPCR,using cyclophilin as housekeeping gene. To study cell viability, cellswere stimulated for 24 or 48 h with 0 (C) 0,4 (IMT0,4), 2,2 (IMT2,2)and 4 ug/ul (IMT4) of IMT504 in DMEM, 20 mM glucose, 0,5% BSA,71 uM βmercaptoethanol. For gene expression, cells were stimulated for 12, 24 and 48 h with the same stimuli. No differences incell viability were observed at the time points studied (ANOVA forrepeated measures: NS; n=5). Expression of Pdx-1 and INS2 wassignificantly increased by 48 h stimulation with IMT504 [ANOVA forrepeated measures: Pdx-1 (A.U.): C=0,47±0,03; IMT0,4=0,40±0,03;IMT2,2=0,52±0,04; IMT4=0,68±0,05 p