Enhancement of diethylstilbestrol induced cytotoxicity by Bcl-2 antisense oligodeoxynucleotides and a glutathione depletor for prostate cancer

Eiji Kikuchi, Jun Nakashima, Yutaka Horiguchi, Mototsugu Oya, Takashi Ohigashi, Masaru Murai

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Purpose: Bcl-2 has been shown to prolong cancer cell survival by blocking apoptosis and have the function of scavenging reactive oxygen species. We assessed the efficacy of a novel strategy that relies on antisense bcl-2 oligodeoxynucleotides as well as a glutathione depletor combined with diethylstilbestrol (DES) for hormone independent prostate cancer. Materials and Methods: The cytotoxic effects of antisense bcl-2 oligodeoxynucleotides and DES on PC-3 cells were determined using spectrophotometry measurement. The expression of Bcl-2 protein was detected by Western blot analysis. Intracellular reactive oxygen species generation was estimated from the amount of intracellular dichlorofluorescein. Apoptosis was determined by the terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate-biotin nick end labeling assay. Results: Antisense bcl-2 oligodeoxynucleotides decreased Bcl-2 protein levels and significantly inhibited PC-3 cell growth in a dose dependent manner. Antisense bcl-2 oligodeoxynucleotides significantly enhanced DES induced cytotoxicity. A significant increase in apoptotic cells was induced by the combination of antisense bcl-2 oligodeoxynucleotides and DES compared with the oligodeoxynucleotides alone. The glutathione depletor buthionine sulfoximine significantly decreased the intracellular concentration of glutathione and augmented DES induced cytotoxicity and reactive oxygen species generation in PC-3 cells. Neither the generation of reactive oxygen species nor the intracellular concentration of glutathione was influenced by antisense bcl-2 oligodeoxynucleotide treatment. Conclusions: Antisense bcl-2 oligodeoxynucleotides significantly enhanced DES induced cytotoxicity in hormone independent prostate cancer cells through the apoptotic pathway independent of augmented reactive oxygen species generation, whereas the glutathione depletor augmented cytotoxicity and reactive oxygen species generation.

Original languageEnglish
Pages (from-to)730-734
Number of pages5
JournalJournal of Urology
Volume169
Issue number2
DOIs
Publication statusPublished - 2003 Feb 1

Fingerprint

Diethylstilbestrol
Glutathione
Prostatic Neoplasms
Reactive Oxygen Species
Hormones
Apoptosis
Buthionine Sulfoximine
DNA Nucleotidylexotransferase
Oligodeoxyribonucleotides
Spectrophotometry
Biotin
oblimersen
Cell Survival
Proteins
Western Blotting
Growth

Keywords

  • Diethylstilbestrol
  • Glutathione
  • Prostate
  • Prostatic neoplasms
  • Reactive oxygen species

ASJC Scopus subject areas

  • Urology

Cite this

Enhancement of diethylstilbestrol induced cytotoxicity by Bcl-2 antisense oligodeoxynucleotides and a glutathione depletor for prostate cancer. / Kikuchi, Eiji; Nakashima, Jun; Horiguchi, Yutaka; Oya, Mototsugu; Ohigashi, Takashi; Murai, Masaru.

In: Journal of Urology, Vol. 169, No. 2, 01.02.2003, p. 730-734.

Research output: Contribution to journalArticle

Kikuchi, Eiji ; Nakashima, Jun ; Horiguchi, Yutaka ; Oya, Mototsugu ; Ohigashi, Takashi ; Murai, Masaru. / Enhancement of diethylstilbestrol induced cytotoxicity by Bcl-2 antisense oligodeoxynucleotides and a glutathione depletor for prostate cancer. In: Journal of Urology. 2003 ; Vol. 169, No. 2. pp. 730-734.
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T1 - Enhancement of diethylstilbestrol induced cytotoxicity by Bcl-2 antisense oligodeoxynucleotides and a glutathione depletor for prostate cancer

AU - Kikuchi, Eiji

AU - Nakashima, Jun

AU - Horiguchi, Yutaka

AU - Oya, Mototsugu

AU - Ohigashi, Takashi

AU - Murai, Masaru

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N2 - Purpose: Bcl-2 has been shown to prolong cancer cell survival by blocking apoptosis and have the function of scavenging reactive oxygen species. We assessed the efficacy of a novel strategy that relies on antisense bcl-2 oligodeoxynucleotides as well as a glutathione depletor combined with diethylstilbestrol (DES) for hormone independent prostate cancer. Materials and Methods: The cytotoxic effects of antisense bcl-2 oligodeoxynucleotides and DES on PC-3 cells were determined using spectrophotometry measurement. The expression of Bcl-2 protein was detected by Western blot analysis. Intracellular reactive oxygen species generation was estimated from the amount of intracellular dichlorofluorescein. Apoptosis was determined by the terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate-biotin nick end labeling assay. Results: Antisense bcl-2 oligodeoxynucleotides decreased Bcl-2 protein levels and significantly inhibited PC-3 cell growth in a dose dependent manner. Antisense bcl-2 oligodeoxynucleotides significantly enhanced DES induced cytotoxicity. A significant increase in apoptotic cells was induced by the combination of antisense bcl-2 oligodeoxynucleotides and DES compared with the oligodeoxynucleotides alone. The glutathione depletor buthionine sulfoximine significantly decreased the intracellular concentration of glutathione and augmented DES induced cytotoxicity and reactive oxygen species generation in PC-3 cells. Neither the generation of reactive oxygen species nor the intracellular concentration of glutathione was influenced by antisense bcl-2 oligodeoxynucleotide treatment. Conclusions: Antisense bcl-2 oligodeoxynucleotides significantly enhanced DES induced cytotoxicity in hormone independent prostate cancer cells through the apoptotic pathway independent of augmented reactive oxygen species generation, whereas the glutathione depletor augmented cytotoxicity and reactive oxygen species generation.

AB - Purpose: Bcl-2 has been shown to prolong cancer cell survival by blocking apoptosis and have the function of scavenging reactive oxygen species. We assessed the efficacy of a novel strategy that relies on antisense bcl-2 oligodeoxynucleotides as well as a glutathione depletor combined with diethylstilbestrol (DES) for hormone independent prostate cancer. Materials and Methods: The cytotoxic effects of antisense bcl-2 oligodeoxynucleotides and DES on PC-3 cells were determined using spectrophotometry measurement. The expression of Bcl-2 protein was detected by Western blot analysis. Intracellular reactive oxygen species generation was estimated from the amount of intracellular dichlorofluorescein. Apoptosis was determined by the terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate-biotin nick end labeling assay. Results: Antisense bcl-2 oligodeoxynucleotides decreased Bcl-2 protein levels and significantly inhibited PC-3 cell growth in a dose dependent manner. Antisense bcl-2 oligodeoxynucleotides significantly enhanced DES induced cytotoxicity. A significant increase in apoptotic cells was induced by the combination of antisense bcl-2 oligodeoxynucleotides and DES compared with the oligodeoxynucleotides alone. The glutathione depletor buthionine sulfoximine significantly decreased the intracellular concentration of glutathione and augmented DES induced cytotoxicity and reactive oxygen species generation in PC-3 cells. Neither the generation of reactive oxygen species nor the intracellular concentration of glutathione was influenced by antisense bcl-2 oligodeoxynucleotide treatment. Conclusions: Antisense bcl-2 oligodeoxynucleotides significantly enhanced DES induced cytotoxicity in hormone independent prostate cancer cells through the apoptotic pathway independent of augmented reactive oxygen species generation, whereas the glutathione depletor augmented cytotoxicity and reactive oxygen species generation.

KW - Diethylstilbestrol

KW - Glutathione

KW - Prostate

KW - Prostatic neoplasms

KW - Reactive oxygen species

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