Antibody to human α-fetoprotein inhibits cell growth of human hepatocellular carcinoma cells by resuscitating the PTEN molecule: in vitro experiments

Ohkawa,Kiyoshi; Asakura,Tadashi; Tsukada,Yutaka; Matsuura,Tomokazu

doi:10.3892/ijo.2017.3982

Antibody to human α-fetoprotein inhibits cell growth of human hepatocellular carcinoma cells by resuscitating the PTEN molecule: in vitro experiments

Authors:
- Kiyoshi Ohkawa
- Tadashi Asakura
- Yutaka Tsukada
- Tomokazu Matsuura
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Affiliations: Stable Isotope Medical Applications Laboratory, Research Center for Medical Science, Jikei University School of Medicine, Minato-ku, Tokyo 105-8461, Japan, Radioisotope Research Facilities, Research Center for Medical Science, Jikei University School of Medicine, Minato-ku, Tokyo 105-8461, Japan, Hachioji Laboratory, SRL Inc., Komiya-cho, Hachioji, Tokyo 192-8535, Japan, Department of Laboratory Medicine, Jikei University School of Medicine, Minato-ku, Tokyo 105-8461, Japan
Published online on: May 3, 2017 https://doi.org/10.3892/ijo.2017.3982
Pages: 2180-2190

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Abstract

It has been proposed that α-fetoprotein (AFP) is a new member of the intracellular signaling molecule family of the phosphoinositide 3-kinase (PI3K)/AKT signaling pathway via interaction with the phosphatase and tensin homolog (PTEN). In this study, the effects of anti-human AFP antibody on the functions of PTEN were examined using an AFP-producing human hepatoma cell line. The antibody caused significant inhibition of cell growth, compared to a normal IgG control, with the accumulation of intracellular immune complexes followed by significant reduction of cytosolic functional AFP. Decrease in the amount of AKT phosphorylated on serine (S) 473 indicated that PI3K/AKT signaling was suppressed in the cells. S380-phosphorylated PTEN increased markedly by the second day after antibody treatment, with slight but significant increase in the PTEN protein level. Since phosphorylation at S380 is critical for PTEN stability, the increase in S380-phosphorylated PTEN indicated maintenance of the number of PTEN molecules and the related potential to control PI3K/AKT signaling. p53 protein (P53) significantly, but slightly increased during antibody treatment, because PTEN expression increased the stability and function of P53 via both molecular interactions. P53 phosphorylated at S20 or at S392 dramatically increased, suggesting an increase in the stability, accumulation and activation of P53. Glucose transporter 1 (GLUT1) increased immediately after antibody treatment, pointing to a deficiency of glucose in the cells. Immunofluorescence cytology revealed that antibody-treatment re-distributed GLUT1 molecules throughout the cytoplasm with a reduction of their patchy localization on the cell surface. This suggested that translocation of GLUT1 depends on the PI3K/AKT pathway, in particular on PTEN expression. Antibody therapy targeted at AFP-producing tumor cells showed an inhibitory effect on the PI3K/AKT pathway via the liberation, restoration and functional stabilization of PTEN. PTEN simultaneously induced both P53 activation and intracellular translocation of GLUT1, since these are closely associated with PTEN.

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