REIC/Dkk-3 stable transfection reduces the malignant phenotype of mouse prostate cancer RM9 cells

Jie Chen, Masami Watanabe, Peng Huang, Masakiyo Sakaguchi, Kazuhiko Ochiai, Yasutomo Nasu, Mamoru Oouchida, Nam Ho Huh, Kenji Shimizu, Yuji Kashiwakura, Haruki Kaku, Hiromi Kumon

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

The reduced expression in immortalized cells (REIC)/Dickkopf (Dkk)-3, a member of the Dkk gene family, is a tumor suppressor in a broad range of cancers. REIC/Dkk-3 transfected stable clones of mouse prostate cancer RM9 cells (RM9-REIC) and the empty vector-transfected control clone cells (RM9-EV) were established. Clones were used to evaluate the anti-cancer effects and a proteomics analysis of REIC/Dkk-3 continuous expression was performed. The RM9-REIC cells show a feeble appearance and the cell membrane shows irregular buds known as blebs. In vitro cell proliferation was significantly suppressed in RM9-REIC clones in comparison to the control. The apoptosis assay was done under standard culture conditions and RM9-REIC showed a higher incidence of apoptosis. The RM9-EV and RM9-REIC cells were orthotopically implanted into a C57BL/ 6 mouse prostate. After 2 weeks, the tumor growth was significantly inhibited in RM9-REIC cells in comparison to the control. Two-dimensional gel electrophoresis was used to examine the modification of protein expression by the gene transfection. The analysis with mass spectrometry disclosed that expression of peroxiredoxin-1, GST-P1, transgelin-2, MRP-L12, ARD, GRP78 and Sorcin were increased and eEF1A-1 and cyclophilin-40 protein were decreased in RM9-REIC cells. Therefore, REIC/Dkk-3 stable transfectants show a reduction of malignancy in mouse prostate cancer RM9 cells in vitro and in vivo. The result of the proteomics analysis might provide important clues to clarify the anti-cancer molecular mechanism of REIC/Dkk-3 gene transfer.

Original languageEnglish
Pages (from-to)789-794
Number of pages6
JournalInternational Journal of Molecular Medicine
Volume24
Issue number6
DOIs
Publication statusPublished - 2009

Fingerprint

Transfection
Prostatic Neoplasms
Phenotype
Clone Cells
Neoplasms
Proteomics
L 012
Apoptosis
Peroxiredoxins
Electrophoresis, Gel, Two-Dimensional
Blister
Inbred C57BL Mouse
Genes
Prostate
Mass Spectrometry
Proteins

Keywords

  • Dickkopf-3
  • Malignant phenotype
  • Prostate cancer
  • Reduced expression in immortalized cells
  • Two-dimensional gelelectrophoresis

ASJC Scopus subject areas

  • Genetics

Cite this

REIC/Dkk-3 stable transfection reduces the malignant phenotype of mouse prostate cancer RM9 cells. / Chen, Jie; Watanabe, Masami; Huang, Peng; Sakaguchi, Masakiyo; Ochiai, Kazuhiko; Nasu, Yasutomo; Oouchida, Mamoru; Huh, Nam Ho; Shimizu, Kenji; Kashiwakura, Yuji; Kaku, Haruki; Kumon, Hiromi.

In: International Journal of Molecular Medicine, Vol. 24, No. 6, 2009, p. 789-794.

Research output: Contribution to journalArticle

Chen, Jie ; Watanabe, Masami ; Huang, Peng ; Sakaguchi, Masakiyo ; Ochiai, Kazuhiko ; Nasu, Yasutomo ; Oouchida, Mamoru ; Huh, Nam Ho ; Shimizu, Kenji ; Kashiwakura, Yuji ; Kaku, Haruki ; Kumon, Hiromi. / REIC/Dkk-3 stable transfection reduces the malignant phenotype of mouse prostate cancer RM9 cells. In: International Journal of Molecular Medicine. 2009 ; Vol. 24, No. 6. pp. 789-794.
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abstract = "The reduced expression in immortalized cells (REIC)/Dickkopf (Dkk)-3, a member of the Dkk gene family, is a tumor suppressor in a broad range of cancers. REIC/Dkk-3 transfected stable clones of mouse prostate cancer RM9 cells (RM9-REIC) and the empty vector-transfected control clone cells (RM9-EV) were established. Clones were used to evaluate the anti-cancer effects and a proteomics analysis of REIC/Dkk-3 continuous expression was performed. The RM9-REIC cells show a feeble appearance and the cell membrane shows irregular buds known as blebs. In vitro cell proliferation was significantly suppressed in RM9-REIC clones in comparison to the control. The apoptosis assay was done under standard culture conditions and RM9-REIC showed a higher incidence of apoptosis. The RM9-EV and RM9-REIC cells were orthotopically implanted into a C57BL/ 6 mouse prostate. After 2 weeks, the tumor growth was significantly inhibited in RM9-REIC cells in comparison to the control. Two-dimensional gel electrophoresis was used to examine the modification of protein expression by the gene transfection. The analysis with mass spectrometry disclosed that expression of peroxiredoxin-1, GST-P1, transgelin-2, MRP-L12, ARD, GRP78 and Sorcin were increased and eEF1A-1 and cyclophilin-40 protein were decreased in RM9-REIC cells. Therefore, REIC/Dkk-3 stable transfectants show a reduction of malignancy in mouse prostate cancer RM9 cells in vitro and in vivo. The result of the proteomics analysis might provide important clues to clarify the anti-cancer molecular mechanism of REIC/Dkk-3 gene transfer.",
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AU - Ochiai, Kazuhiko

AU - Nasu, Yasutomo

AU - Oouchida, Mamoru

AU - Huh, Nam Ho

AU - Shimizu, Kenji

AU - Kashiwakura, Yuji

AU - Kaku, Haruki

AU - Kumon, Hiromi

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