We located that genes, all of that are known to be involved inside the acute inflammatory response, were essentially the most up regulated genes in BMSCs co cultured with leukemia cells. Ingenuity Pathway Evaluation of the differentially expressed genes re vealed that probably the most over represented canonical pathways have been the IL 17 signaling, CD40 signaling and NF?B signal ing pathways. We also compared the micro array data in the distinct time points and we located that the majority of the alterations in the BMSC gene expression profiles occurred inside 4 h. Subsequent, we checked if BMSCs responded differently towards the three various leukemia cell lines. The microarray data were analyzed separately for BMSCs co cultured with all the 3 different leukemia cell lines and we discovered that BMSCs reacted somewhat differently when co cultured with every in the 3 leukemia cell lines.
Working with Partek Genomic Suite, we discovered that the number of differ entially expressed genes in BMSCs co cultured with TF 1, TF 1 and K562 compared with selleck chemicals BMSC mono cultures were 1775, 1375 and 1738 respectively. The genes IL8, CCL2, CXCL1, IL1B and ICAM1 were amongst one of the most up regulated genes in BMSCs co cultured with each TF 1 and K562 although with significantly different fold changes. In contrast, analysis of BMSCs co cultured with TF 1 revealed a unique signature using a mild up regulation of IRF8 and CADHERIN7 and a down regulation of COL3A1.
Ingenuity pathway analysis of your 3 separate sets of BMSC differentially selleck expressed genes revealed that the leading canonical pathways involved were IL 17 signal ing, CD40 signaling and IL 6 signaling in BMSCs co cultured with TF 1 and K562, though Rac signaling, actin cytoskeleton signaling, growth hormone signaling and death receptor signaling have been amongst essentially the most over represented canonical pathways in BMSC co cultured with TF 1. To validate the microarray information, we performed quanti tative RT PCR analysis. The RT PCR results confirmed the higher expression of CCL2, ICAM1, IL8 and IL1B in BMSCs co cultured with leukemia cells compared with BMSC mono cultures. To study the effects of BMSCs on leukemia cells, the gene expression profiles of TF 1, TF 1 and K562 leu kemia cells alone and co cultured with BMSCs were ana lyzed by microarrays. The microarray data were analyzed using Partek Genomic Suite plus the evaluation revealed that 1138, 1119 and 943 genes had been differentially expressed in TF 1, TF 1 and K562 cells co cultured with BMSCs compared with all the respective leukemia cell mono cultures.
Amongst probably the most up regulated genes have been RGS1, FAM69A, Skg1 and SOCSs, even though their fold adjust in expression was 7. Ingenuity pathway analysis from the differentially expressed genes revealed that the most represented canonical pathways were stem cells pluripotency, TGF B signaling and carcinoma signaling.