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Down regulation of N-acetylglucosaminyltransferase V facilitates all-transretinoic acid to induce apoptosis of human hepatocarcinoma cells

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Abstract

After N-acetylglucosaminyltransferase V (GnT-V) activity was down-regulated by the transfection of its antisense cDNA(GnTV-AS), apoptosis of H7721 cells was appeared and the apoptosis induced by 80μM all-transretinoic acid (ATRA) was facilitated, while ATRA itself could not induce apparent apoptosis in mock cells transfected with the vector. In the study of the molecular mechanism of this phenomenon, it was found that GnTV-AS reduced the expressions of anti-apoptotic proteins, such as phosphorylated protein kinase B and phosphorylated Bad as well as Bcl-2 and Bcl-X L, and elevated those of pro-apoptotic proteins, including Bax, full length caspase-3 and its activated fragments as well as anti-oncoprotein p53. In the contrast, ATRA up regulated the expressions of Bax and activated caspase-3 fragments only. After the GnTV-AS transfected cells were treated with ATRA, phosphorylated PKB and Bad were further decreased, while Bax and activated caspase-3 fragment were further increased, leading to the enhanced apoptosis in flow-cytometry analysis when compared with GnTV-AS cells not treated with ATRA. It was speculated that the decreased phospho-Bad resulted from the reduced phospho-PKB and the up regulation of p53 caused the elevated activity of Bax. The increased active caspase-3 was the consequence of the elevated Bax/ Bcl-2(Bcl-X L) activity ratio in the cells.

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Abbreviations

GnT-V:

N-acetylglucosaminyltransferase V

GnTV-AS:

antisense cDNA of N-acetylglucosaminyltransferase V

GlcNAc:

N-acetylglucosamine; Man, mannose;ATRA, all-trans retinoic acid

HPLC:

high performance liquid chromatography

FITC:

fluorescein isothiocyanate; PI, propidium iodide

FCM:

flow-cytometry; FACS, Fluorescence-activated cell spectra

I≖sB:

inhibitor of nuclear factor kappa B (NFκB)

HRP:

horseradish peroxidase; ECL, enhanced chemiluminescence

PKB(Akt):

protein kinase

B; EGTA:

ethylene glycol-bis (â-aminoethyl ester) N,N,N′,N′-tetraacetic acid

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Authors and Affiliations

  1. Key Laboratory of Glycoconjugate Research, Ministry of Health, Department of Biochemistry, Shanghai Medical College, Fudan University, Shanghai, 200032, China

    Peng Guo, Qiu-yan Wang & Hui-Li Chen

  2. Department of Urology, Zhong-Shan Hospital, Fu-Dan University, Shanghai, 200032, China

    Hai-jiao Chen

Authors
  1. Peng Guo
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  2. Hai-jiao Chen
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  3. Qiu-yan Wang
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  4. Hui-Li Chen
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Correspondence to Hui-Li Chen.

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These two persons contributed to this article equally.

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Guo, P., Chen, Hj., Wang, Qy. et al. Down regulation of N-acetylglucosaminyltransferase V facilitates all-transretinoic acid to induce apoptosis of human hepatocarcinoma cells. Mol Cell Biochem 284, 103–110 (2006). https://doi.org/10.1007/s11010-005-9022-5

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  • DOI: https://doi.org/10.1007/s11010-005-9022-5

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