Suppression of α5 gene expression is closely related to the tumorigenic properties of uveal melanoma cell lines

Landreville, Solange; Vigneault, François; Bergeron, Marie-Ève; Leclerc, Suzanne; Gaudreault, Manon; Morcos, Mohib; Mouriaux, Frédéric; Salesse, Christian; Guérin, Sylvain L.

doi:10.1111/j.1755-148x.2011.00869.x

Cited by 17 publications

(22 citation statements)

References 51 publications

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“…27 Isolation and culture of normal uveal melanocytes (UVM) was performed according to the procedure described by Hu et al 28 …”

Section: Cell Culture Of Uveal Melanoma Cell Linesmentioning

confidence: 99%

“…CAT activities were determined and normalized to secreted hGH as described. 27 Each CAT value corresponds to the mean of at least three separate transfections done in triplicate.…”

Section: Transfections and Cat Assaysmentioning

confidence: 99%

“…27,29 The membrane was blotted with a mouse anti-PARG (D8B10; dilution 1:500; Millipore, Billerica, MA), a mouse anti-PARP (C2-10; dilution 1:1,000; a kind gift from Guy Poirier, CHUL Research Center, Québec, Canada), or a mouse anti-ACTIN (dilution 1:40,000; Cedarlane, Burlington, ON, Canada). Antibody (F-20)X (Santa Cruz Biotechnology Inc., Santa Cruz, CA) was used for detection of the ERM TF in the same extracts as for PARP-1 and PARG.…”

Section: Western Blotsmentioning

confidence: 99%

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Altered Expression of the Poly(ADP-Ribosyl)ation Enzymes in Uveal Melanoma and Regulation ofPARGGene Expression by the Transcription Factor ERM

Molloy-Simard

St-Laurent

Vigneault

et al. 2012

Invest. Ophthalmol. Vis. Sci.

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PURPOSE. Poly(ADP-ribosyl)ation is a reversible post-translational modification that requires the contribution of the enzymes poly(ADP-ribose) polymerase-1 (PARP-1) and poly(ADP-ribose) glycohydrolase (PARG). Our study explores expression and activity of PARP-1 and PARG in uveal melanoma cell lines with varying tumorigenic properties.METHODS. Gene profiling on microarrays was conducted using RNA prepared from the uveal melanoma cell lines T97, T98, T108, and T115. The activity of PARP-1 and PARG was monitored by enzymatic assays, whereas their expression was measured by Western blot and PCR. The PARG promoter was analyzed using promoter deletions and site-specific mutagenesis in transfection analyses. The transcription factors binding the PARG promoter were studied by electrophoretic mobility shift assay (EMSA) analyses. Suppression of PARP-1 and PARG expression was performed in T97 and T115 cells by RNAi, and their tumorigenic properties monitored by injections into athymic mice.RESULTS. Expression of PARP-1 was found to vary considerably between uveal melanoma cell lines with distinctive tumorigenic properties in vivo. Sp1 and the ETS protein ERM were shown to bind to the PARG gene promoter to ensure basal transcription in uveal melanoma. Importantly, suppression of PARG gene expression in T97 and T115 cells increased their capacity to form tumors in athymic mice, whereas suppression of PARP-1 significantly reduced or almost entirely abolished tumor formation. CONCLUSIONS.Our results suggest that while overexpression of PARP-1 may confer a proliferative advantage to aggressive uveal melanoma tumors, PARG may, on the other hand, support a tumor suppressor function in vivo. (Invest Ophthalmol Vis Sci.

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“…27 Isolation and culture of normal uveal melanocytes (UVM) was performed according to the procedure described by Hu et al 28 …”

Section: Cell Culture Of Uveal Melanoma Cell Linesmentioning

confidence: 99%

“…CAT activities were determined and normalized to secreted hGH as described. 27 Each CAT value corresponds to the mean of at least three separate transfections done in triplicate.…”

Section: Transfections and Cat Assaysmentioning

confidence: 99%

Section: Western Blotsmentioning

confidence: 99%

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Altered Expression of the Poly(ADP-Ribosyl)ation Enzymes in Uveal Melanoma and Regulation ofPARGGene Expression by the Transcription Factor ERM

Molloy-Simard

St-Laurent

Vigneault

et al. 2012

Invest. Ophthalmol. Vis. Sci.

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“…Data obtained with cell lines derived from four individual donors diagnosed with uveal melanoma show that increased expression of NF1 leads to a more invasive phenotype of uveal melanoma [66]. Aggressiveness of melanomas (as well as of many types of cancers) is related to their ability to interact with the components of the extracellular matrix [67], such as membrane bound receptors and integrins (especially the α5β1 form and its ligand fibronectin).…”

Section: Nf1 In Cancer Development and Progressionmentioning

confidence: 99%

“…The expression of α5 integrin is regulated by NF1 in cooperation with Sp1 and AP1. High expression of NF1 represses the α5 expression and, thus, leads to a more tumorigenic phenotype [66].…”

Section: Nf1 In Cancer Development and Progressionmentioning

confidence: 99%

New insights into the role of NF1 in cancer

Sabová

Kretova²,

Luciaková³

2013

neo

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NF1 proteins are a family of transcription factors that act either as repressors or as activators. Functional studies indicate that NF1 participate in signaling pathways that regulate cell viability, proliferation and differentiation. Participation in regulation of genes important for tumor progression and metastasizing suggests a potential value of NF1 as a prognostic factor for certain types of cancer.

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Contribution of Sp1 to Telomerase Expression and Activity in Skin Keratinocytes Cultured With a Feeder Layer

Bisson

Paquet

Bourget

et al. 2014

Journal Cellular Physiology

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The growth of primary keratinocytes is improved by culturing them with a feeder layer. The aim of this study was to assess whether the feeder layer increases the lifespan of cultured epithelial cells by maintaining or improving telomerase activity and expression. The addition of an irradiated fibroblast feeder layer of either human or mouse origin (i3T3) helped maintain telomerase activity as well as expression of the transcription factor Sp1 in cultured keratinocytes. In contrast, senescence occurred earlier, together with a reduction of Sp1 expression and telomerase activity, in keratinocytes cultured without a feeder layer. Telomerase activity was consistently higher in keratinocytes grown on the three different feeder layers tested relative to cells grown without them. Suppression of Sp1 expression by RNA inhibition (RNAi) reduced both telomerase expression and activity in keratinocytes and also abolished their long-term growth capacity suggesting that Sp1 is a key regulator of both telomerase gene expression and cell cycle progression of primary cultured human skin keratinocytes. The results of the present study therefore suggest that the beneficial influence of the feeder layer relies on its ability to preserve telomerase activity in cultured human keratinocytes through the maintenance of stable levels of Sp1 expression.

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Suppression of α5 gene expression is closely related to the tumorigenic properties of uveal melanoma cell lines

Cited by 17 publications

References 51 publications

Altered Expression of the Poly(ADP-Ribosyl)ation Enzymes in Uveal Melanoma and Regulation ofPARGGene Expression by the Transcription Factor ERM

Altered Expression of the Poly(ADP-Ribosyl)ation Enzymes in Uveal Melanoma and Regulation ofPARGGene Expression by the Transcription Factor ERM

New insights into the role of NF1 in cancer

Contribution of Sp1 to Telomerase Expression and Activity in Skin Keratinocytes Cultured With a Feeder Layer

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