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Title: Involvement of up-regulated Necl-5/Tage4/PVR/CD155 in the loss of contact inhibition in transformed NIH3T3 cells

Abstract

Normal cells show contact inhibition of cell movement and proliferation, but this is lost following transformation. We found that Necl-5, originally identified as a poliovirus receptor and up-regulated in many cancer cells, enhances growth factor-induced cell movement and proliferation. We showed that when cells contact other cells, Necl-5 interacts in trans with nectin-3 and is removed by endocytosis from the cell surface, resulting in a reduction of cell movement and proliferation. We show here that up-regulation of the gene encoding Necl-5 by the oncogene V12-Ki-Ras causes enhanced cell movement and proliferation. Upon cell-cell contact, de novo synthesis of Necl-5 exceeds the rate of Necl-5 endocytosis, eventually resulting in a net increase in the amount of Necl-5 at the cell surface. In addition, expression of the gene encoding nectin-3 is markedly reduced in transformed cells. Thus, up-regulation of Necl-5 following transformation contributes to the loss of contact inhibition in transformed cells.

Authors:
 [1];  [2];  [1];  [1];  [1];  [2];  [3];  [4]
  1. Department of Molecular Biology and Biochemistry, Osaka University Graduate School of Medicine/Faculty of Medicine, Suita 565-0871, Osaka (Japan)
  2. (Japan)
  3. Department of Surgery and Clinical Oncology, Osaka University Graduate School of Medicine/Faculty of Medicine, Suita 565-0871, Osaka (Japan)
  4. Department of Molecular Biology and Biochemistry, Osaka University Graduate School of Medicine/Faculty of Medicine, Suita 565-0871, Osaka (Japan). E-mail: ytakai@molbio.med.osaka-u.ac.jp
Publication Date:
OSTI Identifier:
20857972
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemical and Biophysical Research Communications; Journal Volume: 352; Journal Issue: 4; Other Information: DOI: 10.1016/j.bbrc.2006.11.089; PII: S0006-291X(06)02559-9; Copyright (c) 2006 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ANTIBODIES; BIOSYNTHESIS; CELL PROLIFERATION; GENE REGULATION; GROWTH FACTORS; INHIBITION; MOLECULES; NEOPLASMS; ONCOGENES; POLIO VIRUS; POTASSIUM IODIDES; RECEPTORS; RNA

Citation Formats

Minami, Yukiko, Department of Surgery and Clinical Oncology, Osaka University Graduate School of Medicine/Faculty of Medicine, Suita 565-0871, Osaka, Ikeda, Wataru, Kajita, Mihoko, Fujito, Tsutomu, Department of Surgery and Clinical Oncology, Osaka University Graduate School of Medicine/Faculty of Medicine, Suita 565-0871, Osaka, Monden, Morito, and Takai, Yoshimi. Involvement of up-regulated Necl-5/Tage4/PVR/CD155 in the loss of contact inhibition in transformed NIH3T3 cells. United States: N. p., 2007. Web. doi:10.1016/j.bbrc.2006.11.089.
Minami, Yukiko, Department of Surgery and Clinical Oncology, Osaka University Graduate School of Medicine/Faculty of Medicine, Suita 565-0871, Osaka, Ikeda, Wataru, Kajita, Mihoko, Fujito, Tsutomu, Department of Surgery and Clinical Oncology, Osaka University Graduate School of Medicine/Faculty of Medicine, Suita 565-0871, Osaka, Monden, Morito, & Takai, Yoshimi. Involvement of up-regulated Necl-5/Tage4/PVR/CD155 in the loss of contact inhibition in transformed NIH3T3 cells. United States. doi:10.1016/j.bbrc.2006.11.089.
Minami, Yukiko, Department of Surgery and Clinical Oncology, Osaka University Graduate School of Medicine/Faculty of Medicine, Suita 565-0871, Osaka, Ikeda, Wataru, Kajita, Mihoko, Fujito, Tsutomu, Department of Surgery and Clinical Oncology, Osaka University Graduate School of Medicine/Faculty of Medicine, Suita 565-0871, Osaka, Monden, Morito, and Takai, Yoshimi. Fri . "Involvement of up-regulated Necl-5/Tage4/PVR/CD155 in the loss of contact inhibition in transformed NIH3T3 cells". United States. doi:10.1016/j.bbrc.2006.11.089.
@article{osti_20857972,
title = {Involvement of up-regulated Necl-5/Tage4/PVR/CD155 in the loss of contact inhibition in transformed NIH3T3 cells},
author = {Minami, Yukiko and Department of Surgery and Clinical Oncology, Osaka University Graduate School of Medicine/Faculty of Medicine, Suita 565-0871, Osaka and Ikeda, Wataru and Kajita, Mihoko and Fujito, Tsutomu and Department of Surgery and Clinical Oncology, Osaka University Graduate School of Medicine/Faculty of Medicine, Suita 565-0871, Osaka and Monden, Morito and Takai, Yoshimi},
abstractNote = {Normal cells show contact inhibition of cell movement and proliferation, but this is lost following transformation. We found that Necl-5, originally identified as a poliovirus receptor and up-regulated in many cancer cells, enhances growth factor-induced cell movement and proliferation. We showed that when cells contact other cells, Necl-5 interacts in trans with nectin-3 and is removed by endocytosis from the cell surface, resulting in a reduction of cell movement and proliferation. We show here that up-regulation of the gene encoding Necl-5 by the oncogene V12-Ki-Ras causes enhanced cell movement and proliferation. Upon cell-cell contact, de novo synthesis of Necl-5 exceeds the rate of Necl-5 endocytosis, eventually resulting in a net increase in the amount of Necl-5 at the cell surface. In addition, expression of the gene encoding nectin-3 is markedly reduced in transformed cells. Thus, up-regulation of Necl-5 following transformation contributes to the loss of contact inhibition in transformed cells.},
doi = {10.1016/j.bbrc.2006.11.089},
journal = {Biochemical and Biophysical Research Communications},
number = 4,
volume = 352,
place = {United States},
year = {Fri Jan 26 00:00:00 EST 2007},
month = {Fri Jan 26 00:00:00 EST 2007}
}
  • Previously, we found a significant reduction of progesterone receptor B (PR-B) expression levels in the Ras-mediated NIH3T3 cell transformation, and re-expression of exogenous PR-B eliminated the tumorigenic potential. We hypothesized that this reduction is of biological significance in cell transformation. In the present study, we determined the correlation between PR-B expression and cell cycle progression. In synchronized NIH3T3 cells, we found an increase in PR-B protein and p27 CDK inhibitor levels in the G0/G1 phase and a reduction due to redistribution in the S and G2/M phases. The MEK inhibitor or cAMP stimulation arrested NIH3T3 cells in the G0/G1 phasemore » of the cell cycle. The expression of PR-B and p27 CDK inhibitors was up-regulated by treatment with both the MEK inhibitor and cAMP. Treatment of synchronized cells with a PKA inhibitor in the presence of 1% calf serum resulted in a significant reduction in both PR-B and p27 levels. The decrease in the PR-B levels caused by anti-sense oligomers or siRNA corresponded to the reduction in p27 levels. PR-B overexpression by adenovirus infection induced p27 and suppressed cell growth. Finally, we showed that PR-B modulation involved in the regulation of NIH3T3 cell proliferation was independent of nuclear estrogen receptor (ER) activity but dependent on non-genomic ER activity.« less
  • The RET protooncogene encodes a receptor tyrosine kinase involved in the control differentiation of neural crest derived cells. Point mutations of the RET tyrosine kinase domain were identified among others in 2 distinct genetic disorders, Hirchsprung disease (HSCR) and Multiple Endocrine Neoplasia 2B (MEN 2B). In order to test the biological effect of HSCR and MEN 2B mutations we used a system based on RET-PTC2, a chimeric activated form of the RET protoocogene isolated from a papillary thyroid carcinoma, which shows a detectable transforming activity in NIH3T3 cells and induction of differentiation in PC12 cells. By site-direct mutagenesis we introducedmore » into RET-PTC2 cDNA the mutations at codon 918 (Met{yields}thr, typical of MEN 2B), at codon 765 (Ser{yields}Pro, observed in HSCR) and at codon 897 (Arg{yields}Gln, also observed in HSCR). The former mutation appears to increase the transforming activity of RET-PTC2 in NIH3T3 cells. The latter two mutations abolish the oncogenic activity in NIH3T3 cells as well as its differentiating effect in PC12 cells. These results suggest that RET mutations may cause MEN 2B and HSCR phenotypes through a mechanism of gain or loss of function respectively. Finally, co-transfection experiments of wild-type RET-PTC2 with either HSCR mutation are in progress in order to test the hypothesis of a dominant negative effect in heterozygous state.« less
  • It is well established that DNA replication and ultraviolet-induced DNA repair synthesis in mammalian cells are aphidicolin-sensitive and thus are mediated by one or both of the aphidicolin-sensitive DNA polymerases, ..cap alpha.. and/or delta. Recently, it has been shown that DNA polymerase delta is much more sensitive to inhibition by the nucleotide analogue 2',3'-dideoxythymidine 5'-triphosphate (ddTTP) than DNA polymerase ..cap alpha.. but is less sensitive than DNA polymerase BETA. The authors find that DNA replication and ultraviolet-induced DNA repair synthesis in permeable human fibroblasts are also more sensitive to inhibition by ddTTP than polymerase ..cap alpha.. and less sensitive thanmore » polymerase ..beta... The K/sub i/ for ddTTP of replication is about 40 ..mu..M and that of repair synthesis is about 25 ..mu..M. These are both much less than the K/sub i/ of polymerase ..cap alpha.. (which is greater than 200 ..mu..M but greater than the K/sub i/ of polymerase ..beta.. (which is less than 2 ..mu..M). These data suggest that DNA polymerase delta participates in DNA replication and ultraviolet-induced DNA repair synthesis in human cells.« less
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