Roles of arrest-defective protein 1(225) and hypoxia-inducible factor 1alpha in tumor growth and metastasis.

Abstract

BACKGROUND:

Vascular endothelial growth factor A (VEGFA), a critical mediator of tumor angiogenesis, is a well-characterized target of hypoxia-inducible factor 1 (HIF-1). Murine arrest-defective protein 1A (mARD1A(225)) acetylates HIF-1alpha, triggering its degradation, and thus may play a role in decreased expression of VEGFA.

METHODS:

We generated Apc(Min/+)/mARD1A(225) transgenic mice and quantified growth of intestinal polyps. Human gastric MKN74 and murine melanoma B16F10 cells overexpressing mARD1A(225) were injected into mice, and tumor growth and metastasis were measured. VEGFA expression and microvessel density in tumors were assessed using immunohistochemistry. To evaluate the role of mARD1A(225) acetylation of Lys532 in HIF-1alpha, we injected B16F10-mARD1A(225) cell lines stably expressing mutant HIF-1alpha/K532R into mice and measured metastasis. All statistical tests were two-sided, and P values less than .05 were considered statistically significant.

RESULTS:

Apc(Min/+)/mARD1A(225) transgenic mice (n = 25) had statistically significantly fewer intestinal polyps than Apc(Min/+) mice (n = 21) (number of intestinal polyps per mouse: Apc(Min/+) mice vs Apc(Min/+)/mARD1A(225) transgenic mice, mean = 83.4 vs 38.0 polyps, difference = 45.4 polyps, 95% confidence interval [CI] = 41.8 to 48.6; P < .001). The growth and metastases of transplanted tumors were also statistically significantly reduced in mice injected with mARD1A(225)-overexpressing cells than in mice injected with control cells (P < .01). Moreover, overexpression of mARD1A(225) decreased VEGFA expression and microvessel density in tumor xenografts (P < .04) and Apc(Min/+) intestinal polyps (P = .001). Mutation of lysine 532 of HIF-1alpha in B16F10-mARD1A(225) cells prevented HIF-1alpha degradation and inhibited the antimetastatic effect of mARD1A(225) (P < .001).

CONCLUSION:

mARD1A(225) may be a novel upstream target that blocks VEGFA expression and tumor-related angiogenesis.

• Authors: Mi-Ni Lee, Shi-Nai Lee, Se-Hee Kim, Bora Kim, Bo-Kyung Jung, Ji Hae Seo, Ji-Hyeon Park, Jae-Hoon Choi, Sun Hee Yim, Mi-Ran Lee, Jong-Gil Park, Ji-Young Yoo, Jeong Hun Kim, Seung-Taek Lee, Hwan-Mook Kim, Sandra Ryeom, Kyu-Won Kim, Goo Taeg Oh
• Journal: Journal of the National Cancer Institute (J Natl Cancer Inst) Vol. 102 Issue 6 Pg. 426-42 (Mar 17 2010) ISSN: 1460-2105 [Electronic] United States
• PMID: 20194889 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• HIF1A protein, human
• Hypoxia-Inducible Factor 1, alpha Subunit
• Vascular Endothelial Growth Factor A
• Acetyltransferases
• N-Terminal Acetyltransferase A
• Naa10 protein, mouse
• N-Terminal Acetyltransferase E
• Lysine

Topics

• Acetylation
• Acetyltransferases (metabolism)
• Animals
• Blotting, Western
• Cell Line, Tumor
• Down-Regulation
• Enzyme-Linked Immunosorbent Assay
• Female
• Gene Expression Regulation, Neoplastic
• Humans
• Hypoxia-Inducible Factor 1, alpha Subunit (genetics, metabolism)
• Immunohistochemistry
• Immunoprecipitation
• Intestinal Polyps (metabolism, pathology)
• Lung Neoplasms (metabolism, secondary)
• Lysine
• Male
• Mice
• Mice, Inbred BALB C
• Mice, Inbred C57BL
• Mice, Nude
• Mice, Transgenic
• Microcirculation
• Mutation
• N-Terminal Acetyltransferase A
• N-Terminal Acetyltransferase E
• Neovascularization, Pathologic (metabolism, prevention & control)
• Reverse Transcriptase Polymerase Chain Reaction
• Up-Regulation
• Vascular Endothelial Growth Factor A (analysis)