Abstract
The modulation of GnT-V activity by signaling molecules in PI-3-K/PKB pathway in human hepatocarcinoma cell line 7721 was studied. GnT-V activity was determined after the transfection of sense or antisense cDNA of PKB into the cells, as well as the addition of activators, specific inhibitors, and the antibodies to the enzyme assay system or culture medium. It was found that the basal activity of GnT-V was up regulated by the sense and down regulated by the antisense cDNA of PKB transfected into 7721 cells. GnT-V was activated by PIP2, PIP3 or GTPγ[S] added to the assay system, and the activation of PIP2 or GTPγ[S] was abolished by LY2940002, a specific inhibitor of PI-3-K, but the activation of PIP3 was not attenuated by LY2940002. In addition, GnT-V activity in cultured parental or H-ras transfected cells was inhibited by the antibody against PKB or PI-3-K. These findings demonstrated the involvement of PI-3-K/PKB signaling pathway in the regulation of GnT-V. Moreover, ET18-OCH3, an inhibitor of Raf translocation and PI-PLC enzyme, which produces the activator of PKC, as well as the antibodies against Raf-1 or MEK also inhibited GnT-V activity in the parental and H-ras transfected cells. The inhibitory rates, however, were less in the transfected cells than those in the parental cells. These results reveal that in parental and H-ras transfected 7721 cells, the basal activity of GnT-V is also regulated by the Ras/Raf-1/MEK/MAPK cascade in addition to PI-3-K/PKB signaling pathway. The significance of these two pathways in the regulation of GnT-V and their relations to the activation of PKC previously reported by our laboratory (Ju TZ et al., 1995 Glyconjugate J 12, 767–772) was discussed.
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References
Brockhausen I, Narasimhan S, and Schachter H, The biosynthesis of highly branched N-glycans. Studies on the sequential pathway and functional role of N-acetylglycosaminyltransferase I,II,III,IV,V and VI, Biochimie 70, 1521–33 (1988).
Pierce M, Buckhaults P, Chen L, and Fregien N, Regulation of N-acetylgluco-saminyltransferase V and Asn-linked oligosaccharide b(1,6) branching by a growth factor signaling pathway and effects on cell adhesion and metastatic potential, Glycoconj J 14, 623–30 (1997).
Nan BC, Shao DM, Chen HL, Huang Y, Gu JX, Zhang YB, and Wu ZG, Alteration of N-acetylglucosaminyltransferase in pancreatic cancer, Glycoconj J 15, 1033–7 (1998).
Ju TZ, Chen HL, Gu JX, and Qin H, Regulation of N-acetylglucosaminyl transferase V by protein kinase, Glycoconj J 12, 767–72 (1995).
Babara MM, and Julian D, PKB/Akt: connecting phosphoinositides 3-kinase to cell survival and beyond, TIBS 22, 355–8 (1997).
Hemming BA, PtdIns (3,4,5) P3 gets its message across, Science 277, 534–6 (1997).
Franke TF, Kaplan DR, and Cantley LC, PI3K: Downstream AKTion blocks apoptosis, Cell 88, 435–7 (1997).
Lee SA, and Holz RW, Protein phosphorylation and secretion in digitonin-permeabilized adrenal chromaffin cells. Effects of micromolar Ca2+, phorbol esters, and diacylglycerol, J Biol Chem 261, 17089–98 (1986).
Ausybel FM, Brent R, Kinston RE, Moore DD, Seidman JG, Smith JG, and Struhl K, Short Protocols in Molecular Biology (3rd ed.) pp 9.11–9.12, <q>4.22–4.25 (1995).
Lowry OH, Rosebrough NJ, Farr AL, and Randall RJ, Protein measurement with the Folinphenol-reagent, J Biol Chem 193, 265–71 (1951).
Marone M, Scambia G, Ferrandina G, Ghiannitelli C, Benedetti-Panici P, Iacovella S, Leone A, and Mancuso S, Nm23 expression in endometrial and cervical cancer inverse correlation with lymph node involvement and myometrial invasion, Brit J Cancer 74, 1063–8 (1996).
Coffer P, and Woodgett JR, Molecular cloning and characterization of a novel putative protein-serine kinase related to cAMPdependent and protein kinase C families, Eur J Biochem 201, 475–81 (1991).
Standaert M, Bandyopadhyay G, Galloway L, Ono Y, Mukai H, and Farese R, Comparative effects of GTPgamaS and insulin on the activation of Rho, phosphatidylinositol 3-kinase, and protein kinase N in rat adipocytes, J Biol Chem 273, 7470–7 (1998).
Cohen P, Alessi DR, and Cross DAE, PDK1, tone of the missing links in insulin transduction, FEBS letter 410, 3–10 (1997).
Zhou X, Lu X, Richard C, Xiong W, Litchfield DW, Bittman R, and Authur G, 1-O-Octadecyl-2-methyl-glycerophosphocholine inhibits the transduction of growth signals via the MAPK cascade in cultured MCF-7 cells, J Clin Invest 98, 937–44 (1996).
Powis G, Seewald MJ, Gratas C, Melder D, Riebow J, Modest, EJ, Selective inhibition of phosphatidylinositol phospholipase C by cytotoxic ether lipid analoques, Cancer Res 52, 2835–40 (1992).
Wang QH, Zhou DP, Jiang SM, Shen ZH, and Gu JX, Effects of epidermal growth factor and insulin on the activity of Nacetylglucosaminyltransferase V, Biochem J 324, 543–5 (1998).
Nakamura SI, Phosphatidylcholine hydrolysis and protein C activation for intracellular signaling network, J Lipid Mediat Cell Signalling 14, 197–202 (1996).
Xia BY, Guo HB, and Chen HL, Dual regulation of tyrosine protein kinase and protein kinase C on N-acetylglucosaminyltransferase V, Acta Biochim Biophys Sinica 30, 421–31 (1998).
Delcommenne M, Tan C, Gray V, Rue L, Woodkett J, and Dedhar S, Phosphoonositide-3-OH kinase-dependent regulation of glyco-Modification of GnT-V activity by PI-3-K and PKB 321 gen synthetase kinase 3 and protein kinase B/AKT by the integrin-linked kinase, Proc Natl Acad Sci USA 95, 11211–6 (1998).
Rubio I, Rodriguez Viciana P, Dowmward J, and Wetzker R, Interaction of Ras with phosphoinositide 3-kinase gama, Biochem J 326, 891–5 (1997).
Wymann MP, and Pirola L, Structure and function of phosphoinositide 3-kinase. Biochim Biophys Acta 1436, 127–50 (1998).
Guo HB, Jiang AL, Ju TZ, and Chen HL, Opposing changes in N-acetylglucosaminyl-transferase Vand III during the cell cycle and all-trans retinoic acid treatment of hepatocarcinoma cell line, Biochim Biophys Acta 1495, 297–307 (2000).
Guo HB, Liu F, Zhao JH, and Chen HL, Down regulation of Nacetylglucosaminyl transferase V by tumorigenesis and metastasis suppressor gene and its relation to metastatic potential of human hepatocarcinoma cells, J Cellular Biochem 79, 370–385 (2000).
Lopez-Ilasaca M, Crespo P, Pellici PG, Gitkind JS, and Wetzker R, Linkage of G protein-coupled receptor to the MAPK signalling pathwa through PI 3-kinase gamma, Science 275, 394–7 (1997).
Lopez-Ilasaca M, Gitkind JS, and Wetzker R, Phosphoinositide 3-kinase Îû is a mediater of GÎôÎû-dependent Jun kinase activation, J Biol Chem 273, 2505–8 (1998).
Marais R, Light Y, Mason C, Paterson H, Olson MF, and Marshall CJ, Requirement of Ras-GTP-Raf complexes for activation of Raf-1 by protein kinase C, Science 280, 109–12 (1998).
Sable CL, Filippa N, Hemming B, and Van Obberghen E, cAMP stimulates protein kinase B in a Wortmanin-insensitive manner, FEBS Lett 409, 253–7 (1997).
Dumont JE, Jauniaux JC, and Roger PP, The cyclic AMP mediated stimulation of celll proliferation, TIBS 14, 67–71 (1989).
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Guo, Hb., Shen, Zh., Huang, Cx. et al. Modulation of the basal activity of N-acetylglucosaminyltransferase V by phosphatidylinositol-3-kinase/protein kinase B signaling pathway in human hepatocarcinoma cells. Glycoconj J 17, 315–322 (2000). https://doi.org/10.1023/A:1007177806496
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DOI: https://doi.org/10.1023/A:1007177806496