Establishment and characterization of human ocular melanoma cell lines

Aubert, C; Rougé, Françoise; Reillaudou, M; Metge, P

doi:10.1002/ijc.2910540513

Cited by 35 publications

(18 citation statements)

References 12 publications

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“…Despite the difficulties in culturing human UM cells, we achieved a success rate similar to that described by other authors [Aubert et al, 1993]. We found that primary cell cultures derived from UM have a slow growing rate and need between 6 and 10 days to double their number.…”

Section: Discussion Um Primary Cell Cultures and Cell Cyclesupporting

confidence: 82%

Abnormal cell cycle regulation in primary human uveal melanoma cultures

Pardo

Piñeiro

Fuente

et al. 2004

J of Cellular Biochemistry

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Uveal malignant melanoma is the most frequent primary intraocular tumor in adult humans. The cellular events leading to neoplasic transformation of normal uveal melanocytes are not well known when compared to other cancers. In this study, we investigated the role of G1 and G1/S regulatory proteins of the cell cycle in human uveal melanoma (UM) primary cell cultures, since these proteins are common targets in tumor development. Further, freshly established and characterized tumor cells are a better model for in vitro studies when compared to cell lines established long ago. Human primary cell cultures from eight different UM were established, as well as one primary culture from rhesus uveal normal melanocytes (UNM). Primary human UM cultures were characterized by a low establishment and growing rate. From four successful cultures, three showed a high expression of cyclin D1, cyclin E, p16NK4A, and p27KIP1 with no variations in cyclin A, cyclin-dependent kinase 2 (CDK2), and CDK4. Interestingly, in one of the cultured tumors, tumor suppressor protein retinoblastoma (Rb) did not bind E2F despite the fact that Rb was found in its hypophosphorylated form. No mutations in either RB1 or the Rb-binding pocket of E2F-1 were detected. Furthermore, we identified seven proteins co-immunoprecipitating with Rb in this tumor, including Lamin A/C and six proteins not previously reported to bind Rb: Hsc70, high mobility group protein 1 (HMG-1), hnRPN, glyceraldehyde 3 phosphate dehydrogenase (G3PDH), EF-1, and EF-2. Our results indicate that the overexpression of cyclins D1/E and CDKIs p16 and p27, together with a deregulation of the Rb/E2F pathway, may be implicated in the development of human UM.

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Section: Discussion Um Primary Cell Cultures and Cell Cyclesupporting

confidence: 82%

Abnormal cell cycle regulation in primary human uveal melanoma cultures

Pardo

Piñeiro

Fuente

et al. 2004

J of Cellular Biochemistry

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“…The melanoma cell lines JPC 298 (Aubert et al, 1993), A375 (Giard et al, 1973), Mewo (Bean, 1975), P1 (kindly provided by Dr Dummer, Zurich), SK-Mel28 (Carey et al, 1976), HT144 (Ahrens et al, 2001), Bro (Lockshin et al, 1985) were maintained in RPMI 1640 with stable glutamine supplemented with penicillin (400 U/ml), streptomycin (50 mg/ml), ascorbic acid (50 mg/ml) and 10% fetal calf serum (Seromed s , Biochrom KG; Berlin, Germany), in a humidified atmosphere of 5% CO 2 at 371C.…”

Section: Cell Lines and Culture Conditionsmentioning

confidence: 99%

Interaction of human Thy-1 (CD 90) with the integrin αvβ3 (CD51/CD61): an important mechanism mediating melanoma cell adhesion to activated endothelium

et al. 2005

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The expression of the avb3 integrin (CD51/CD61) on human melanoma cells has been shown to be associated most closely with tumor progression and metastases formation in melanoma. Here, we demonstrated a specific interaction of the avb3 integrin on melanoma cells with the human Thy-1, an inducible cell adhesion molecule expressed on the cell surface of activated endothelial cells (EC). The interaction was shown by the binding of purified Thy-1 protein to a V b 3 transfected cells, to avb3-expressing melanoma cells and to purified a V b 3 integrin. Moreover, melanoma cells adhere specifically to Thy-1 transfectants via avb3 on melanoma cells showing the functional relevance of this interaction for cell adhesion. Finally, the importance of the avb3/Thy-1 interaction for the adhesion of melanoma cells to the activated endothelium was confirmed under static and flow conditions by the inhibition of melanoma cell adhesion to and transmigration across activated EC by blocking the avb3/Thy-1 interaction. In conclusion, we have identified a new pair of adhesion molecules Thy-1 and avb3 mediating the interaction of melanoma cells and activated EC. These data explain at least in part the high tumorigenicity of avb3-expressing melanoma cells and the association of avb3-positive melanoma cells with a high risk of metastasis and poor prognosis.

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“…In contrast to short-term cultivation, performing studies by using cell lines has the advantage of stable and reproducible experimental conditions caused by regular growth patterns of the cells. For this reason, several attempts for the establishment of cell lines derived from primary uveal melanomas were made in the past [4,5,6,7,8]. Compared to other malignancies, uveal melanomas are rare and difficult to cultivate.…”

Section: Introductionmentioning

confidence: 99%

Novel Cell Lines Derived by Long-Term Culture of Primary Uveal Melanomas

et al. 2009

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Aims: Uveal melanomas with or without monosomy 3 have different metastatic potentials. Therefore, it would be of great experimental importance to obtain cell lines of both groups – established and characterized under the same conditions. Methods: Long-term culture of biopsies derived from untreated primary uveal melanomas was performed, and the status of chromosome 3 in the tumour was determined. Characterization by immunocytochemistry and in vitro assays for cell migration, vasculogenic cord formation and proliferation were performed. Results: Establishment of cell lines succeeded in 6 out of 128 uveal melanomas. No histopathological feature was predictive for cultivation success. Two out of 4 cell lines derived from tumours with monosomy 3 were invasive in vitro. Both cell lines with disomy 3 showed higher proliferation levels (p = 0.002). Conclusions: The in vitro tendency for invasion did not correlate with the proliferative behaviour of uveal melanoma cell lines, but was partly associated with monosomy of chromosome 3.

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Establishment and characterization of human ocular melanoma cell lines

Cited by 35 publications

References 12 publications

Abnormal cell cycle regulation in primary human uveal melanoma cultures

Abnormal cell cycle regulation in primary human uveal melanoma cultures

Interaction of human Thy-1 (CD 90) with the integrin αvβ3 (CD51/CD61): an important mechanism mediating melanoma cell adhesion to activated endothelium

Novel Cell Lines Derived by Long-Term Culture of Primary Uveal Melanomas

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