Karyotype instability and anchorage-independent growth in telomerase-immortalized fibroblasts from two centenarian individuals

Mondello, Chiara; Chiesa, Massimo; Rebuzzini, Paola; Zongaro, Samantha; Verri, Annalisa; Colombo, Tina; Giulotto, Elena; D’Incalci, Maurizio; Franceschi, Claudio; Nuzzo, Fiorella

doi:10.1016/s0006-291x(03)01484-0

Cited by 50 publications

(70 citation statements)

References 46 publications

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“…Mondello et al (29) observed karyotype abnormalities (both numerical and structural) and anchorage-independent growth in soft agar of fibroblasts that were isolated from two centenarian patients and subsequently infected with hTERT by using a retroviral vector. However, no anchorage-independent growth was observed before PD 60, and no hTERT cell populations that were injected into immunodeficient mice produced tumors after 3 months, despite inoculation at PD107 (29).…”

Section: Discussionmentioning

confidence: 99%

“…However, no anchorage-independent growth was observed before PD 60, and no hTERT cell populations that were injected into immunodeficient mice produced tumors after 3 months, despite inoculation at PD107 (29). Milyavsky et al (37) showed that a subpopulation of hTERT-immortalized fibroblasts gradually developed inactivation of p16 INK4A and associated loss of contact inhibition as well as increased susceptibility to transformation by H-Ras.…”

Section: Discussionmentioning

confidence: 99%

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Relevance and safety of telomerase for human tissue engineering

Klinger

Blum

Hearn

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

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Tissue engineering holds the promise of replacing damaged or diseased tissues and organs. The use of autologous donor cells is often not feasible because of the limited replicative lifespan of cells, particularly those derived from elderly patients. Proliferative arrest can be overcome by the ectopic expression of telomerase via human telomerase reverse transcriptase (hTERT) gene transfection. To study the efficacy and safety of this potentially valuable technology, we used differentiated vascular smooth muscle cells (SMC) and vascular tissue engineering as a model system. Although we previously demonstrated that vessels engineered with telomerase-expressing SMC had improved mechanics over those grown with control cells, it is critical to assess the phenotypic impact of telomerase expression in donor cells, because telomerase upregulation is observed in >95% of human malignancies. To study the impact of telomerase in tissue engineering, expression of hTERT was retrovirally induced in SMC from eight elderly patients and one young donor. In hTERT SMC, significant lifespan extension beyond that of control was achieved without population doubling time acceleration. Karyotype changes were seen in both control and hTERT SMC but were not clonal nor representative of cancerous change. hTERT cells also failed to show evidence of neoplastic transformation in functional assays of tumorigenicity. In addition, the impact of donor age on cellular behavior, particularly the synthetic capability of SMC, was not affected by hTERT expression. Hence, this tissue engineering model system highlights the impact of donor age on cellular synthetic function that appears to be independent of lifespan extension by hTERT.tumorigenicity ͉ smooth muscle cells ͉ vascular graft

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Relevance and safety of telomerase for human tissue engineering

Klinger

Blum

Hearn

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

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“…The cen3tel cellular system has been previously described (11,12). Briefly, it was derived from primary cen3 fibroblasts by infection with a human telomerase reverse transcriptase (hTERT)-containing retrovirus (13). Telomerase-expressing cells were propagated in culture to population doubling (PD) ~1,000, and a gradual acquisition of the neoplastic phenotype was observed (11,12).…”

Section: Methodsmentioning

confidence: 99%

Snail levels control the migration mechanism of mesenchymal tumor cells

et al. 2016

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Abstract. Cancer cells use two major types of movement:Mesenchymal, which is typical of cells of mesenchymal origin and depends on matrix metalloproteinase (MMP) activity, and amoeboid, which is characteristic of cells with a rounded shape and relies on the activity of Rho-associated kinase (ROCK). The present authors previously demonstrated that, during neoplastic transformation, telomerase-immortalized human fibroblasts (cen3tel cells) acquired a ROCK-dependent/MMP independent mechanism of invasion, mediated by the downregulation of the ROCK cellular inhibitor Round (Rnd)3/RhoE. In the present study, cen3tel transformation was also demonstrated to be paralleled by downregulation of Snail, a major determinant of the mesenchymal movement. To test whether Snail levels could determine the type of movement adopted by mesenchymal tumor cells, Snail was ectopically expressed in tumorigenic cells. It was observed that ectopic Snail did not increase the levels of typical mesenchymal markers, but induced cells to adopt an MMP-dependent mechanism of invasion. In cells expressing ectopic Snail, invasion became sensitive to the MMP inhibitor Ro 28-2653 and insensitive to the ROCK inhibitor Y27632, suggesting that, once induced by Snail, the mesenchymal movement prevails over the amoeboid one. Snail-expressing cells had a more aggressive behavior in vivo, and exhibited increased tumor growth rate and metastatic ability. These results confirm the high plasticity of cancer cells, which can adopt different types of movement in response to changes in the expression of specific genes. Furthermore, the present findings indicate that Rnd3 and Snail are possible regulators of the type of invasion mechanism adopted by mesenchymal tumor cells.

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“…The cen3tel telomerase immortalized cell line was obtained from primary cen3 fibroblasts, derived from a centenarian individual, by infection with an hTERTcontaining retrovirus (8). Cen3tel cells were used at different steps of propagation (up to around PD 1000) reflecting different phases of transformation (7)(8)(9).…”

Section: Methodsmentioning

confidence: 99%

“…During culture propagation, immortalized cells, named cen3tel, underwent neoplastic transformation, they became able to induce tumors when subcutaneously injected into nude mice and then acquired metastatic capacity (7)(8)(9). Cen3tel cells at the early stages of tumorigenicity formed pleomorphic sarcomas in nude mice, cells at later stages induced tumors with a hemangioperycitoma-like vascular pattern, analogous to that observed in synovial sarcomas.…”

Section: Introductionmentioning

confidence: 98%

Relocalization of cell adhesion molecules during neoplastic transformation of human fibroblasts

Belgiovine

Chiodi²,

Mondello

2011

Int J Oncol

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Abstract. Studying neoplastic transformation of telomerase immortalized human fibroblasts (cen3tel), we found that the transition from normal to tumorigenic cells was associated with the loss of growth contact inhibition, the acquisition of an epithelial-like morphology and a change in actin organization, from stress fibers to cortical bundles. We show here that these variations were paralleled by an increase in N-cadherin expression and relocalization of different adhesion molecules, such as N-cadherin, α-catenin, p-120 and β-catenin. These proteins presented a clear membrane localization in tumorigenic cells compared to a more diffuse, cytoplasmic distribution in primary fibroblasts and non-tumorigenic immortalized cells, suggesting that tumorigenic cells could form strong cell-cell contacts and cell contacts did not induce growth inhibition. The epithelial-like appearance of tumorigenic cells did not reflect a mesenchymal-epithelial transition; in fact, cen3tel cells expressed vimentin and did not express cytokeratins at all transformation stages. Moreover, they did not express epithelial proteins such as occluding and claudin-1. In contrast, ZO-1 showed higher levels and a more defined membrane localization in tumorigenic cells compared to non-tumorigenic cells; this confirms its role in adherens junction formation in mesenchymal cells and is in agreement with the strong cellcell contact formation by neoplastically transformed cells. Finally, we found α-catenin and ZO-1 nuclear localization in non-transformed cells, suggestive of possible additional roles of these proteins besides cell junction formation.

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Karyotype instability and anchorage-independent growth in telomerase-immortalized fibroblasts from two centenarian individuals

Cited by 50 publications

References 46 publications

Relevance and safety of telomerase for human tissue engineering

Relevance and safety of telomerase for human tissue engineering

Snail levels control the migration mechanism of mesenchymal tumor cells

Relocalization of cell adhesion molecules during neoplastic transformation of human fibroblasts

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