Acceleration of Gastric Tumorigenesis Through MKRN1-Mediated Posttranslational Regulation of p14ARF

Aram Ko, Ji Young Shin, Jinho Seo, Kang Duck Lee, Eun Woo Lee, Min Sik Lee, Han Woong Lee, Il Ju Choi, Jin Sook Jeong, Kyung Hee Chun, Jaewhan Song

Research output: Contribution to journalArticle

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Abstract

BackgroundWe investigated whether Makorin ring finger protein 1 (MKRN1), an E3 ligase, affects p14ARF-associated cellular senescence and tumorigenesis by posttranslational modification in gastric tumorigenesis.MethodsA link between MKRN1 and ARF was examined in MKRN1 null mouse embryonic fibroblasts (MEFs) and in human fibroblasts and gastric cancer cells by silencing MKRN1 using small interfering RNA (siRNA) and short hairpin RNA (shRNA). Ubiquitination and proteasomal degradation assays were used to assess p14ARF degradation associated with MKRN1. MKRN1 and p14ARF expression levels were analyzed with immunohistochemistry in malignant and normal tissues from gastric cancer patients and with χ2 tests. The tumor growth of gastric cancer cells stably expressing MKRN1 shRNA, p14ARF shRNA, or both was examined in mouse xenograft models (n 4-6) and analyzed with unpaired t tests. All statistical tests were two-sided.ResultsMKRN1 knockout MEFs exhibited premature senescence and growth retardation with increased p19ARF protein expression. Similar results were obtained for human fibroblasts or gastric cancer cell lines by MKRN1 knockdown. Biochemical analyses confirmed that MKRN1 targets p14ARF for ubiquitination and subsequent proteasome-dependent degradation. A statistically significant association was shown between MKRN1 overexpression and p14ARF underexpression (P . 016). Xenograft analyses using p53-functional AGS or -dysfunctional SNU601 cells displayed statistically significant tumor growth retardation by silencing MKRN1, which was reversed under depletion of p14ARF (AGS cells, MKRN1 knockdown tumors vs MKRN1 and p14ARF knockdown tumors: 164.6 vs 464.8mm3, difference 300.2mm3, 95% CI 189.1 to 411.3mm3, P <. 001).ConclusionsWe demonstrated that MKRN1 functions as a novel E3 ligase of p14ARF and that it potentially regulates cellular senescence and tumorigenesis in gastric cancer.

Original languageEnglish
Pages (from-to)1660-1672
Number of pages13
JournalJournal of the National Cancer Institute
Volume104
Issue number21
DOIs
Publication statusPublished - 2012 Nov 7

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Tumor Suppressor Protein p14ARF
Stomach
Carcinogenesis
Stomach Neoplasms
Small Interfering RNA
Fibroblasts
Ubiquitin-Protein Ligases
Cell Aging
Ubiquitination
Makorin ring finger protein 1
Heterografts
Neoplasms
Growth
Proteasome Endopeptidase Complex
Post Translational Protein Processing

All Science Journal Classification (ASJC) codes

  • Oncology
  • Cancer Research

Cite this

Ko, Aram ; Shin, Ji Young ; Seo, Jinho ; Lee, Kang Duck ; Lee, Eun Woo ; Lee, Min Sik ; Lee, Han Woong ; Choi, Il Ju ; Jeong, Jin Sook ; Chun, Kyung Hee ; Song, Jaewhan. / Acceleration of Gastric Tumorigenesis Through MKRN1-Mediated Posttranslational Regulation of p14ARF. In: Journal of the National Cancer Institute. 2012 ; Vol. 104, No. 21. pp. 1660-1672.
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title = "Acceleration of Gastric Tumorigenesis Through MKRN1-Mediated Posttranslational Regulation of p14ARF",
abstract = "BackgroundWe investigated whether Makorin ring finger protein 1 (MKRN1), an E3 ligase, affects p14ARF-associated cellular senescence and tumorigenesis by posttranslational modification in gastric tumorigenesis.MethodsA link between MKRN1 and ARF was examined in MKRN1 null mouse embryonic fibroblasts (MEFs) and in human fibroblasts and gastric cancer cells by silencing MKRN1 using small interfering RNA (siRNA) and short hairpin RNA (shRNA). Ubiquitination and proteasomal degradation assays were used to assess p14ARF degradation associated with MKRN1. MKRN1 and p14ARF expression levels were analyzed with immunohistochemistry in malignant and normal tissues from gastric cancer patients and with χ2 tests. The tumor growth of gastric cancer cells stably expressing MKRN1 shRNA, p14ARF shRNA, or both was examined in mouse xenograft models (n 4-6) and analyzed with unpaired t tests. All statistical tests were two-sided.ResultsMKRN1 knockout MEFs exhibited premature senescence and growth retardation with increased p19ARF protein expression. Similar results were obtained for human fibroblasts or gastric cancer cell lines by MKRN1 knockdown. Biochemical analyses confirmed that MKRN1 targets p14ARF for ubiquitination and subsequent proteasome-dependent degradation. A statistically significant association was shown between MKRN1 overexpression and p14ARF underexpression (P . 016). Xenograft analyses using p53-functional AGS or -dysfunctional SNU601 cells displayed statistically significant tumor growth retardation by silencing MKRN1, which was reversed under depletion of p14ARF (AGS cells, MKRN1 knockdown tumors vs MKRN1 and p14ARF knockdown tumors: 164.6 vs 464.8mm3, difference 300.2mm3, 95{\%} CI 189.1 to 411.3mm3, P <. 001).ConclusionsWe demonstrated that MKRN1 functions as a novel E3 ligase of p14ARF and that it potentially regulates cellular senescence and tumorigenesis in gastric cancer.",
author = "Aram Ko and Shin, {Ji Young} and Jinho Seo and Lee, {Kang Duck} and Lee, {Eun Woo} and Lee, {Min Sik} and Lee, {Han Woong} and Choi, {Il Ju} and Jeong, {Jin Sook} and Chun, {Kyung Hee} and Jaewhan Song",
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Acceleration of Gastric Tumorigenesis Through MKRN1-Mediated Posttranslational Regulation of p14ARF. / Ko, Aram; Shin, Ji Young; Seo, Jinho; Lee, Kang Duck; Lee, Eun Woo; Lee, Min Sik; Lee, Han Woong; Choi, Il Ju; Jeong, Jin Sook; Chun, Kyung Hee; Song, Jaewhan.

In: Journal of the National Cancer Institute, Vol. 104, No. 21, 07.11.2012, p. 1660-1672.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Acceleration of Gastric Tumorigenesis Through MKRN1-Mediated Posttranslational Regulation of p14ARF

AU - Ko, Aram

AU - Shin, Ji Young

AU - Seo, Jinho

AU - Lee, Kang Duck

AU - Lee, Eun Woo

AU - Lee, Min Sik

AU - Lee, Han Woong

AU - Choi, Il Ju

AU - Jeong, Jin Sook

AU - Chun, Kyung Hee

AU - Song, Jaewhan

PY - 2012/11/7

Y1 - 2012/11/7

N2 - BackgroundWe investigated whether Makorin ring finger protein 1 (MKRN1), an E3 ligase, affects p14ARF-associated cellular senescence and tumorigenesis by posttranslational modification in gastric tumorigenesis.MethodsA link between MKRN1 and ARF was examined in MKRN1 null mouse embryonic fibroblasts (MEFs) and in human fibroblasts and gastric cancer cells by silencing MKRN1 using small interfering RNA (siRNA) and short hairpin RNA (shRNA). Ubiquitination and proteasomal degradation assays were used to assess p14ARF degradation associated with MKRN1. MKRN1 and p14ARF expression levels were analyzed with immunohistochemistry in malignant and normal tissues from gastric cancer patients and with χ2 tests. The tumor growth of gastric cancer cells stably expressing MKRN1 shRNA, p14ARF shRNA, or both was examined in mouse xenograft models (n 4-6) and analyzed with unpaired t tests. All statistical tests were two-sided.ResultsMKRN1 knockout MEFs exhibited premature senescence and growth retardation with increased p19ARF protein expression. Similar results were obtained for human fibroblasts or gastric cancer cell lines by MKRN1 knockdown. Biochemical analyses confirmed that MKRN1 targets p14ARF for ubiquitination and subsequent proteasome-dependent degradation. A statistically significant association was shown between MKRN1 overexpression and p14ARF underexpression (P . 016). Xenograft analyses using p53-functional AGS or -dysfunctional SNU601 cells displayed statistically significant tumor growth retardation by silencing MKRN1, which was reversed under depletion of p14ARF (AGS cells, MKRN1 knockdown tumors vs MKRN1 and p14ARF knockdown tumors: 164.6 vs 464.8mm3, difference 300.2mm3, 95% CI 189.1 to 411.3mm3, P <. 001).ConclusionsWe demonstrated that MKRN1 functions as a novel E3 ligase of p14ARF and that it potentially regulates cellular senescence and tumorigenesis in gastric cancer.

AB - BackgroundWe investigated whether Makorin ring finger protein 1 (MKRN1), an E3 ligase, affects p14ARF-associated cellular senescence and tumorigenesis by posttranslational modification in gastric tumorigenesis.MethodsA link between MKRN1 and ARF was examined in MKRN1 null mouse embryonic fibroblasts (MEFs) and in human fibroblasts and gastric cancer cells by silencing MKRN1 using small interfering RNA (siRNA) and short hairpin RNA (shRNA). Ubiquitination and proteasomal degradation assays were used to assess p14ARF degradation associated with MKRN1. MKRN1 and p14ARF expression levels were analyzed with immunohistochemistry in malignant and normal tissues from gastric cancer patients and with χ2 tests. The tumor growth of gastric cancer cells stably expressing MKRN1 shRNA, p14ARF shRNA, or both was examined in mouse xenograft models (n 4-6) and analyzed with unpaired t tests. All statistical tests were two-sided.ResultsMKRN1 knockout MEFs exhibited premature senescence and growth retardation with increased p19ARF protein expression. Similar results were obtained for human fibroblasts or gastric cancer cell lines by MKRN1 knockdown. Biochemical analyses confirmed that MKRN1 targets p14ARF for ubiquitination and subsequent proteasome-dependent degradation. A statistically significant association was shown between MKRN1 overexpression and p14ARF underexpression (P . 016). Xenograft analyses using p53-functional AGS or -dysfunctional SNU601 cells displayed statistically significant tumor growth retardation by silencing MKRN1, which was reversed under depletion of p14ARF (AGS cells, MKRN1 knockdown tumors vs MKRN1 and p14ARF knockdown tumors: 164.6 vs 464.8mm3, difference 300.2mm3, 95% CI 189.1 to 411.3mm3, P <. 001).ConclusionsWe demonstrated that MKRN1 functions as a novel E3 ligase of p14ARF and that it potentially regulates cellular senescence and tumorigenesis in gastric cancer.

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