A specific increase in inositol 1,4,5-trisphosphate 3-kinase B expression upon differentiation of human embryonic stem cells

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• 作者：Catherine Hoofd^a ; Fabienne Devreker^a ; ^{fdevreke@ulb.ac.be} ; Laurence Deneubourg^b ; Sandrine Deleu^a ; Thi Mai Uyen Nguyen^a ; Karen Sermon^c ; Yvon Englert^a ; Christophe Erneux^b ; ^{cerneux@ulb.ac.be}
• 关键词：hECCs ; human embryonal carcinoma cells ; hESCs ; human embryonic stem cells ; mESCs ; mouse embryonic stem cells ; MEF ; mouse embryonic fibroblasts ; FBS ; fetal bovine serum ; rFGF ; recombinant fibroblast growth factor ; GCTs ; germ cell tumours ; [3H] Ins ; tritiat
• 刊名：Cellular Signalling
• 出版年：2012
• 期刊代码：42_08986568
• 类别：bio
• 出版时间：July, 2012
• 卷：24
• 期：7
• 页码：1461-1470
• 文件大小：1461 K

摘要

Human embryonic stem cells (hESCs) are of great hope for regenerative medicine due to their dual pluripotency and self-renewal properties. We report a comparison of inositol phosphate (InsP_s) production in undifferentiated, differentiated hESCs and in two cancer cell lines, Ntera2 cells, a human embryonal carcinoma cell (hECC) line and HeLa cells. To evaluate the potential impact of InsP_s in differentiation, hESCs were spontaneously differentiated in culture for two weeks. The distribution of the different InsP_s was affected upon differentiation: the level of highly phosphorylated InsP_s was decreased. In contrast, the total level of phosphoinositides (PI) was increased. Using real time quantitative PCR (qPCR), the mRNA expression of several enzymes of the metabolism of InsP_s was determined: a specific increase in inositol 1,4,5-trisphosphate 3-kinase A and B (ITPKA and ITPKB) was observed upon hESCs spontaneous differentiation. Ins(1,4,5)P₃ 3-kinase activity, undetectable in undifferentiated hESCs, increased upon differentiation. The same observation was made by Western blotting using an antibody directed against human ITPKB. This is the first report showing the potential implication of soluble InsP_s in hESCs and possible function of isoenzymes of the inositol trisphosphate 3-kinase family in differentiation.