Upregulation of the expression of Fas antigen and Fas ligand in a human submandibular gland ductal cell line by okadaic acid

Yasuhiro Morimoto, Hiroyuki Morimoto, Hirohiko Okamura, Kimiko Nomiyama, Nobuaki Nakamuta, Shigeru Kobayashi, Shinji Kito, Takeshi Ohba, Tatsuji Haneji

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Fas receptor is a member of a superfamily of receptors characterized by cysteine-rich motifs in the extracellular domain of the molecule. Binding of Fas ligand to the receptor leads to receptor activation and the induction of intracellular signals that result in apoptotic cell death. In the present study, the expression of mRNA and proteins of Fas receptor and Fas ligand were examined in human submandibular gland ductal (HSG) cells treated with okadaic acid by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunoblot analysis. Six hundred and eighty-two bp of the PCR product of Fas receptor mRNA was detected in HSG cells and a protein with an estimated molecular weight of 58,000 was expressed in HSG cells. Treatment of HSG cells with an agonistic anti-Fas monoclonal antibody resulted in death of HSG cells, indicating that the functional Fas receptor protein is expressed in HSG cells. Fas receptor protein expression stimulated by okadaic acid was elevated in a dose- and time-dependent manner, with maximal expression at 20 nM and 48 h treatment. Fas ligand mRNA was also detected constitutively in HSG cells by RT-PCR. Okadaic acid stimulated the expression of Fas ligand protein in HSG cells in a time-dependent manner, while the expression of the ligand was low in untreated HSG cells. The molecular weight of Fas ligand was estimated as 68,000. An antagonistic anti-Fas ligand monoclonal antibody prevented okadaic acid-induced death in HSG cells in a dose-dependent fashion as determined by WST-1 assay. The results indicate that the expression of Fas receptor and ligand is regulated by protein phosphatase(s) sensitive to okadaic acid and is involved in okadaic acid-induced apoptosis in HSG cells. The results also suggest that the Fas receptor-ligand system might regulate apoptosis in HSG cells. (C) 200 Elsevier Science Ltd.

Original languageEnglish
Pages (from-to)657-666
Number of pages10
JournalArchives of Oral Biology
Volume45
Issue number8
DOIs
Publication statusPublished - Aug 1 2000
Externally publishedYes

Fingerprint

CD95 Antigens
Okadaic Acid
Fas Ligand Protein
Submandibular Gland
Up-Regulation
Cell Line
anti-Fas monoclonal antibody
Reverse Transcriptase Polymerase Chain Reaction
Messenger RNA
Proteins
Molecular Weight
Apoptosis
Phosphoprotein Phosphatases
Cysteine

Keywords

  • Apoptosis
  • Fas antigen
  • Fas ligand
  • HSG cells
  • Okadaic acid

ASJC Scopus subject areas

  • Dentistry(all)

Cite this

Upregulation of the expression of Fas antigen and Fas ligand in a human submandibular gland ductal cell line by okadaic acid. / Morimoto, Yasuhiro; Morimoto, Hiroyuki; Okamura, Hirohiko; Nomiyama, Kimiko; Nakamuta, Nobuaki; Kobayashi, Shigeru; Kito, Shinji; Ohba, Takeshi; Haneji, Tatsuji.

In: Archives of Oral Biology, Vol. 45, No. 8, 01.08.2000, p. 657-666.

Research output: Contribution to journalArticle

Morimoto, Yasuhiro ; Morimoto, Hiroyuki ; Okamura, Hirohiko ; Nomiyama, Kimiko ; Nakamuta, Nobuaki ; Kobayashi, Shigeru ; Kito, Shinji ; Ohba, Takeshi ; Haneji, Tatsuji. / Upregulation of the expression of Fas antigen and Fas ligand in a human submandibular gland ductal cell line by okadaic acid. In: Archives of Oral Biology. 2000 ; Vol. 45, No. 8. pp. 657-666.
@article{c14a0b6a42fd44f6b435fc7262525725,
title = "Upregulation of the expression of Fas antigen and Fas ligand in a human submandibular gland ductal cell line by okadaic acid",
abstract = "Fas receptor is a member of a superfamily of receptors characterized by cysteine-rich motifs in the extracellular domain of the molecule. Binding of Fas ligand to the receptor leads to receptor activation and the induction of intracellular signals that result in apoptotic cell death. In the present study, the expression of mRNA and proteins of Fas receptor and Fas ligand were examined in human submandibular gland ductal (HSG) cells treated with okadaic acid by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunoblot analysis. Six hundred and eighty-two bp of the PCR product of Fas receptor mRNA was detected in HSG cells and a protein with an estimated molecular weight of 58,000 was expressed in HSG cells. Treatment of HSG cells with an agonistic anti-Fas monoclonal antibody resulted in death of HSG cells, indicating that the functional Fas receptor protein is expressed in HSG cells. Fas receptor protein expression stimulated by okadaic acid was elevated in a dose- and time-dependent manner, with maximal expression at 20 nM and 48 h treatment. Fas ligand mRNA was also detected constitutively in HSG cells by RT-PCR. Okadaic acid stimulated the expression of Fas ligand protein in HSG cells in a time-dependent manner, while the expression of the ligand was low in untreated HSG cells. The molecular weight of Fas ligand was estimated as 68,000. An antagonistic anti-Fas ligand monoclonal antibody prevented okadaic acid-induced death in HSG cells in a dose-dependent fashion as determined by WST-1 assay. The results indicate that the expression of Fas receptor and ligand is regulated by protein phosphatase(s) sensitive to okadaic acid and is involved in okadaic acid-induced apoptosis in HSG cells. The results also suggest that the Fas receptor-ligand system might regulate apoptosis in HSG cells. (C) 200 Elsevier Science Ltd.",
keywords = "Apoptosis, Fas antigen, Fas ligand, HSG cells, Okadaic acid",
author = "Yasuhiro Morimoto and Hiroyuki Morimoto and Hirohiko Okamura and Kimiko Nomiyama and Nobuaki Nakamuta and Shigeru Kobayashi and Shinji Kito and Takeshi Ohba and Tatsuji Haneji",
year = "2000",
month = "8",
day = "1",
doi = "10.1016/S0003-9969(00)00038-8",
language = "English",
volume = "45",
pages = "657--666",
journal = "Archives of Oral Biology",
issn = "0003-9969",
publisher = "Elsevier Limited",
number = "8",

}

TY - JOUR

T1 - Upregulation of the expression of Fas antigen and Fas ligand in a human submandibular gland ductal cell line by okadaic acid

AU - Morimoto, Yasuhiro

AU - Morimoto, Hiroyuki

AU - Okamura, Hirohiko

AU - Nomiyama, Kimiko

AU - Nakamuta, Nobuaki

AU - Kobayashi, Shigeru

AU - Kito, Shinji

AU - Ohba, Takeshi

AU - Haneji, Tatsuji

PY - 2000/8/1

Y1 - 2000/8/1

N2 - Fas receptor is a member of a superfamily of receptors characterized by cysteine-rich motifs in the extracellular domain of the molecule. Binding of Fas ligand to the receptor leads to receptor activation and the induction of intracellular signals that result in apoptotic cell death. In the present study, the expression of mRNA and proteins of Fas receptor and Fas ligand were examined in human submandibular gland ductal (HSG) cells treated with okadaic acid by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunoblot analysis. Six hundred and eighty-two bp of the PCR product of Fas receptor mRNA was detected in HSG cells and a protein with an estimated molecular weight of 58,000 was expressed in HSG cells. Treatment of HSG cells with an agonistic anti-Fas monoclonal antibody resulted in death of HSG cells, indicating that the functional Fas receptor protein is expressed in HSG cells. Fas receptor protein expression stimulated by okadaic acid was elevated in a dose- and time-dependent manner, with maximal expression at 20 nM and 48 h treatment. Fas ligand mRNA was also detected constitutively in HSG cells by RT-PCR. Okadaic acid stimulated the expression of Fas ligand protein in HSG cells in a time-dependent manner, while the expression of the ligand was low in untreated HSG cells. The molecular weight of Fas ligand was estimated as 68,000. An antagonistic anti-Fas ligand monoclonal antibody prevented okadaic acid-induced death in HSG cells in a dose-dependent fashion as determined by WST-1 assay. The results indicate that the expression of Fas receptor and ligand is regulated by protein phosphatase(s) sensitive to okadaic acid and is involved in okadaic acid-induced apoptosis in HSG cells. The results also suggest that the Fas receptor-ligand system might regulate apoptosis in HSG cells. (C) 200 Elsevier Science Ltd.

AB - Fas receptor is a member of a superfamily of receptors characterized by cysteine-rich motifs in the extracellular domain of the molecule. Binding of Fas ligand to the receptor leads to receptor activation and the induction of intracellular signals that result in apoptotic cell death. In the present study, the expression of mRNA and proteins of Fas receptor and Fas ligand were examined in human submandibular gland ductal (HSG) cells treated with okadaic acid by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunoblot analysis. Six hundred and eighty-two bp of the PCR product of Fas receptor mRNA was detected in HSG cells and a protein with an estimated molecular weight of 58,000 was expressed in HSG cells. Treatment of HSG cells with an agonistic anti-Fas monoclonal antibody resulted in death of HSG cells, indicating that the functional Fas receptor protein is expressed in HSG cells. Fas receptor protein expression stimulated by okadaic acid was elevated in a dose- and time-dependent manner, with maximal expression at 20 nM and 48 h treatment. Fas ligand mRNA was also detected constitutively in HSG cells by RT-PCR. Okadaic acid stimulated the expression of Fas ligand protein in HSG cells in a time-dependent manner, while the expression of the ligand was low in untreated HSG cells. The molecular weight of Fas ligand was estimated as 68,000. An antagonistic anti-Fas ligand monoclonal antibody prevented okadaic acid-induced death in HSG cells in a dose-dependent fashion as determined by WST-1 assay. The results indicate that the expression of Fas receptor and ligand is regulated by protein phosphatase(s) sensitive to okadaic acid and is involved in okadaic acid-induced apoptosis in HSG cells. The results also suggest that the Fas receptor-ligand system might regulate apoptosis in HSG cells. (C) 200 Elsevier Science Ltd.

KW - Apoptosis

KW - Fas antigen

KW - Fas ligand

KW - HSG cells

KW - Okadaic acid

UR - http://www.scopus.com/inward/record.url?scp=0033917650&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033917650&partnerID=8YFLogxK

U2 - 10.1016/S0003-9969(00)00038-8

DO - 10.1016/S0003-9969(00)00038-8

M3 - Article

C2 - 10869477

AN - SCOPUS:0033917650

VL - 45

SP - 657

EP - 666

JO - Archives of Oral Biology

JF - Archives of Oral Biology

SN - 0003-9969

IS - 8

ER -