Most considerably differentially phosphorylated peptides in these seven pathways showed greater phosphorylation levels in osteosarcoma cell lines (Figure four), indicating that kinases affect phosphorylation of molecules playing a role in genomic stability and cell cycle progression.PI3K/Akt and AMPK signaling in osteosarcomathe most substantially impacted pathway in osteosarcoma cells (Figure 5) and also the AMPK pathway as second most significantly affected pathway (Further file 9). Specifically, molecules directly downstream of Akt kinases showed larger phosphorylation in osteosarcoma than in MSCs, although molecules downstream of AMPK showed reduce phosphorylation levels. As these results indicate that Akt signaling is active in osteosarcoma and may be driving its higher proliferative capacity, we set out to pharmacologically inhibit Akt using the compound MK-2206.MK-2206 inhibits proliferation of U-2 OS and HOS, but not of 143BUnsupervised pathway analysis on the kinome profiling benefits returned the IPA pathway PI3K/Akt signaling asWe inhibited osteosarcoma and control cells for 120 h with MK-2206, an allosteric inhibitor of all three Akt members of the family. Inhibition of your good handle leukemiaKuijjer et al. BMC Medical Genomics 2014, 7:four http://biomedcentral/1755-8794/7/Page six ofColor Key-1 -0.0.Row Z-ScoreRAF1_253_265QRQRSTSTPNVHM VASP_271_283LARRRKATQVGEK E1A_ADE05_212_224AILRRPTSPVSRE PRKDC_2618_2630TRTQEGSLSARWP P53_12_24PPLSQETFSDLWK KIF2C_105_118_S106GEGLRSRSTRMSTVS CDN1A_139_151GRKRRQTSMTDFY PP2AB_297_309EPHVTRRTPDYFL RBL2_655_667GLGRSITSPTTLY BAD_112_124RELRRMSDEFVDS PLEK_106_118GQKFARKSTRRSI FOXO3_25_37QSRPRSCTWPLQR NEK2_172_184FAKTFVGTPYYMS IKKB_686_698QLMSQPSTASNSL IKKB_173_185_C179ALDQGSLATSFVGT KCNA3_461_473EELRKARSNSTLS DESP_2842_2854RSGSRRGSFDATG FRAP_2443_2455RTRTDSYSAGQSV LMNA_192_204DAENRLQTMKEEL KPCB_19_31_A25SRFARKGSLRQKNV NCF1_296_308RGAPPRRSSIRNA KAP2_92_104SRFNRRVSVCAET SCN7A_898_910KNGCRRGSSLGQI REL_260_272KMQLRRPSDQEVS RS6_228_240IAKRRRLSSLRAS NMDZ1_890_902SFKRRRSSKDTST ADRB2_338_350ELLCLRRSSLKAY KPB1_1011_1023QVEFRRLSISAES ART_025_CXGLRRWSLGGLRRWSLGLRRWSLGGLRRWSL CAC1C_1974_1986ASLGRRASFHLEC CREB1_126_138EILSRRPSYRKIL LIPS_944_956GFHPRRSSQGATQ VTNC_390_402NQNSRRPSRATWL PTK6_436_448ALRERLSSFTSYE CSF1R_701_713NIHLEKKYVRRDS NCF1_321_333QDAYRRNSVRFLQ KAP3_107_119NRFTRRASVCAEA MYPC3_268_280LSAFRRTSLAGGG PTN12_32_44FMRLRRLSTKYRT F263_454_466NPLMRRNSVTPLA GBRB2_427_439SRLRRRASQLKIT CFTR_730_742EPLERRLSLVPDS GPSM2_394_406PKLGRRHSMENME CFTR_761_773LQARRRQSVLNLM GRIK2_708_720FMSSRRQSVLVKS EPB42_241_253LLNKRRGSVPILR TY3H_65_77FIGRRQSLIEDAR KCNA6_504_516ANRERRPSYLPTP VASP_150_162EHIERRVSNAGGPMSCMSC143BFigure 3 Supervised clustering of kinome profiling results.3-DL-Cpa-OH Data Sheet Supervised clustering on all 49 significantly differentially phosphorylated peptides identified by the comparison of two osteosarcoma cell lines with two MSC cultures.3-Methylcyclopentane-1-carboxylic acid In stock Peptides are sorted on logFC, from lower phosphorylation to higher phosphorylation in osteosarcoma cell lines.PMID:23724934 Orange: higher phosphorylation levels, blue: lower phosphorylation levels.cell line NALM-6, and of osteosarcoma cell line U-2 OS with MK-2206 was dose-dependent, with IC50s of 0.38 M and two.five M, and maximal responses of 94 and 71 , respectively (Figure six). 143B did not show any response at concentrations below five M. Mainly because 143B exhibits an oncogenic KRAS transformation [28], we assessed MK-2206 specificity on the parental cell line of 143B, HOS, which has not been KRAS transformed. HOS indee.