Immortalization with telomerase of the Nestin-positive cells of the human pancreas

Lee, K. M.; Nguyen, Christine L.; Ulrich, Alexis; Pour, Parviz M.; Ouellette, Michel

doi:10.1016/s0006-291x(03)00086-x

Cited by 132 publications

(147 citation statements)

References 27 publications

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“…Primary human cells that have thus far been immortalized by the use of hTERT alone include fibroblasts, retinal pigmented epithelial cells, endothelial cells, myometrial cells, oesophageal squamous cells, mammary epithelial cells, keratinocytes, osteoblasts, and Nestin-positive cells of the pancreas (Bodnar et al 1998;Yang et al 1999;Ouellette et al 2000;Ramirez et al 2001;Yudoh et al 2001;Condon et al 2002;Herbert et al 2002;Lee et al 2003;Morales et al 2003). Because primary human cells are typically hard to transfect, exogenous hTERT is generally transferred through the use of retroviral vectors.…”

Section: Introductionmentioning

confidence: 99%

Use of exogenous hTERT to immortalize primary human cells

2004

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A major obstacle to the immortalization of primary human cells and the establishment of human cell lines is telomere-controlled senescence. Telomere-controlled senescence is caused by the shortening of telomeres that occurs each time somatic human cells divide. The enzyme telomerase can prevent the erosion of telomeres and block the onset of telomere-controlled senescence, but its expression is restricted to the early stages of embryonic development, and in the adult, to rare cells of the blood, skin and digestive track. However, we and others have shown that the transfer of an exogenous hTERT cDNA, encoding the catalytic subunit of human telomerase, can be used to prevent telomere shortening, overcome telomerecontrolled senescence, and immortalize primary human cells. Most importantly, hTERT alone can immortalize cells without causing cancer-associated changes or altering phenotypic properties. Primary human cells that have so far been established by the forced expression of hTERT alone include fibroblasts, retinal pigmented epithelial cells, endothelial cells, oesophageal squamous cells, mammary epithelial cells, keratinocytes, osteoblasts, and Nestin-positive cells of the pancreas. In this article, we discuss the use of hTERT to immortalize of human cells, the properties of hTERT-immortalized cells, and their applications to cancer research and tissue engineering.Abbreviations: hTERT -human Telomerase Reverse Transcriptase; hTR -human Telomerase RNA.

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Section: Introductionmentioning

confidence: 99%

Use of exogenous hTERT to immortalize primary human cells

2004

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“…We expect that the attachment of an activator domain to our ZFP-TF targeting the hTERT promoter region could also lead to interesting results, as telomerase activity and telomere lengths are closely related to cell aging. There are many trials using gene insertion (37,38). Telomerase activation using ZFPs should be unique in that a dormant gene is activated.…”

Section: Discussionmentioning

confidence: 99%

Repression of Human Telomerase Reverse Transcriptase Using Artificial Zinc Finger Transcription Factors

Sohn

Yeh

Choi

et al. 2010

Molecular Cancer Research

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Telomerase activation is a key step in the development of human cancers. Expression of the catalytic subunit, human telomerase reverse transcriptase (hTERT), represents the limiting factor for telomerase activity. In this study, we have used artificial zinc finger protein (ZFP) transcription factors (TF) to repress the expression of hTERT in human cancer cell lines at the transcriptional level. We have constructed four-fingered ZFPs derived from the human genome which binds 12-bp recognition sequences within the promoter of the hTERT gene and fused them with a KRAB repressor domain to create a potent transcriptional repressor. Luciferase activity was decreased by >80% in all of the transcriptional repressors with luciferase reporter assay. When they were transfected into the telomerase-positive HEK293 cell line, a decrease of mRNA level and telomerase activity together with shortening of telomere length was observed. Actual growth of HEK293 cells was also inhibited by transfection of artificial ZFP-TFs. The repression was maintained for 100 days of culture. The repression of telomerase expression by artificial ZFP-TFs targeting the promoter region of the hTERT presents a new promising strategy for inhibiting the growth of human cancer cells. Mol Cancer Res; 8(2); 246-53.

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“…No authentication was performed on the HuPT3 cell line by the authors. The immortalized human pancreatic duct-derived (HPNE) cells were described previously (30, 31). …”

Section: Methodsmentioning

confidence: 99%

Response to MLN8237 in Pancreatic Cancer Is Not Dependent on RalA Phosphorylation

Neel

Stratford

Shinde

et al. 2014

Molecular Cancer Therapeutics

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The high prevalence of KRAS mutations and importance of the RalGEF-Ral pathway downstream of activated K-Ras in pancreatic ductal adenocarcinoma (PDAC) emphasize the importance of identifying novel methods by which to therapeutically target these pathways. It was recently demonstrated that phosphorylation of RalA S194 by Aurora A kinase is critical for PDAC tumorigenesis. We sought to evaluate the Aurora A kinase-selective inhibitor MLN8237 as a potential indirect anti-RalA targeted therapy for PDAC. We utilized a site-specific phospho-S194 RalA antibody and determined that RalA S194 phosphorylation levels were elevated in a subset of PDAC cell lines and human tumors relative to unmatched normal controls. Effects of MLN8237 on anchorage-independent growth in PDAC cell lines and growth of patient-derived xenografts (PDX) were variable, with a subset of cell lines and PDX showing sensitivity. Surprisingly, RalA S194 phosphorylation levels in PDAC cell lines or PDX tumors did not correlate with MLN8237 responsiveness. However, we identified Ki67 as a possible early predictive biomarker for response to MLN8237 in PDAC. These results indicate that MLN8237 treatment may be effective for a subset of PDAC patients independent of RalA S194 phosphorylation. Ki67 may be an effective pharmacodynamic biomarker to identify response early in the course of treatment.

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Immortalization with telomerase of the Nestin-positive cells of the human pancreas

Cited by 132 publications

References 27 publications

Use of exogenous hTERT to immortalize primary human cells

Use of exogenous hTERT to immortalize primary human cells

Repression of Human Telomerase Reverse Transcriptase Using Artificial Zinc Finger Transcription Factors

Response to MLN8237 in Pancreatic Cancer Is Not Dependent on RalA Phosphorylation

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