ING4 regulates JWA in angiogenesis and their prognostic value in melanoma patients

Abstract

Background:We previously showed that inhibitor of growth family member 4 (ING4) inhibits melanoma angiogenesis, and JWA suppresses the metastasis of melanoma cells. As angiogenesis is essential for tumour metastasis, further investigation of the function of ING4 and JWA in melanoma angiogenesis is needed, and their prognostic value are of great interest.Methods:Western blot, tube-formation assays and luciferase assays were used to investigate the correlation between ING4 and JWA in melanoma angiogenesis. JWA and integrin-linked kinase (ILK) expression was determined on a tissue microarray constructed from 175 biopsies.Results:ING4 promoted JWA expression by activating JWA promoter. Furthermore, the regulation of growth and tube formation of endothelial cells by ING4 was partially JWA dependent. Also, ING4 inhibited the ILK-induced angiogenesis signalling pathway via JWA. Moreover, reduced JWA, or increased ILK, expression was closely associated with 5-year disease-specific survival of melanoma patients (P=0.001 and 0.007, respectively). There was also a positive correlation between ING4 and JWA yet a negative correlation between ING4 and ILK. Importantly, their concomitant expressions were significantly related to 5-year survival of melanoma patients (P=0.002 and 0.003, respectively).Conclusion:JWA has an important role in ING4-regulated melanoma angiogenesis, and ING4/JWA/ILK are promising prognostic markers and may be used as anti-angiogenic therapeutic targets for melanoma.

The Emerging Role of Inhibitor of Growth 4 as a Tumor Suppressor in Multiple Human Cancers

“…Hou et al also found that ING4 could act as an E3 ubiquitin ligase to induce ubiquitination and degradation of p65, which is an additional mechanism to link ING4 to NF-κB [80]. In addition, ING4 was the upstream regulator of JWA, activating JWA promoter, inhibiting ILK and downstream NF-κB/IL-6/STAT3/VEGF signaling pathways, thus promoting melanoma angiogenesis [81]. In melanoma, ING4 was induced by BRMS1 (Breast cancer metastasis suppressor 1), a metastasis suppressor, and regulated angiogenesis through inhibiting NF-κB activity and IL-6 level [82].…”

132 Eyes absent gene (EYA1) is a pathogenic driver and a therapeutic target for melanoma

“…The selection of melanoma tissue blocks and construction of tumor tissue microarrays have been described previously [6,7,12,[33][34][35][36][37]. The formalinfixed, paraffin embedded archival biopsies of benign melanocytic nevi, melanoma in situ, invasive primary melanoma and metastatic melanoma were obtained from the 1990 to 2009 archives from the Department of Pathology at Vancouver General Hospital, Vancouver, Canada.…”

“…The clinicopathological data was available for all melanoma cases. EYA1 expression was analyzed by immunohistochemistry on paraffin-embedded tissue microarrays as described previously [6,7,12,[33][34][35][36][37]. Briefly, de-paraffinized 4μm tissue sections were treated 30 minutes in 0.1M sodium citrate (PH6.0) at 95˚C for antigen retrieval.…”

“…A previously described [6,7,12,[33][34][35][36][37]] a 4-point scoring system was used to determine intensity of EYA1 staining. Scoring was performed by three independent scorers, including a dermatopathologist, without access to clinico-pathological information of the sections.…”

“…Previous research from our group [5][6][7][8][9][10][11][12][13] and others [3,[14][15][16][17][18][19][20] have revealed numerous genes significantly upregulated in melanoma biopsies compared with benign nevi, and many of these have been shown to contribute to melanoma pathogenesis in in vitro and in vivo experimental systems [21,22]. The genes implicated in melanoma development have been demonstrated to be involved in regulation of cell cycle progression, extracellular matrix remodeling, migration, apoptosis resistance, and many other pathways.…”