Evaluation of Telomerase in Canine Mammary Tissues by Immunohistochemical Analysis and a Polymerase Chain Reaction-Based Enzyme-Linked Immunosorbent Assay

Panarese, Serena; Brunetti, Barbara; Sarli, Giuseppe

doi:10.1177/104063870601800407

Cited by 8 publications

(5 citation statements)

References 49 publications

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“…Telomerase expression, which is assessed by h-TERT IHC, was displayed in expected locations, mainly nucleolar and nuclear locations, as previously reported, and also was rarely found in cytoplasmic locations [9]. Strong nucleolar staining may be explained as the presence of the telomerase holoenzyme, which is assembled within the nucleolus.…”

Section: Discussionsupporting

confidence: 71%

Immunohistochemical expression of h-telomerase reverse transcriptase in canine and feline meningiomas

et al. 2007

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Telomere length maintenance is regarded as a fundamental step in tumorigenesis, as most human brain tumors, including meningiomas, stabilize the ends of their chromosomes using telomerase. This investigation represents an introduction to telomerase expression in canine and feline meningiomas. Twenty-five archived cases (14 dogs and 11 cats) were immunohistochemically tested for human-telomerase reverse transcriptase (h-TERT), scored, and quantified; furthermore, mitoses were counted on sections stained with a modified toluidine blue. The h-TERT antibody immunolabelled the nucleus and nucleolus of meningeal neoplastic cells, with an intensity ranging from mild to strong and a speckled distribution; a significantly higher expression in cats was noted, while no significant association between h-TERT immunolabelling and sex or histotype was evident in dogs or cats. The telomerase enzyme represents a fundamental parameter of potential malignant transformation, which may occur independently of the signal to proliferate, thereby supplying the cells with unlimited growth capabilities. Telomerase expression could be a prognostic indicator independent of the kinetic parameters, although this should be evaluated using a larger dataset with available clinical information.

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Section: Discussionsupporting

confidence: 71%

Immunohistochemical expression of h-telomerase reverse transcriptase in canine and feline meningiomas

et al. 2007

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“…It was thus suggested that reduced phosphorylation either renders hTERT unable to bind to its nuclear translocators or forces the protein to move from the nucleus to the cytoplasm. The fact that the majority of the testicular tumours stained strongly and gave the typical localisation pattern of hTERT (nuclear, cytoplasmic and both nuclear and cytoplasmic) using the NCLhTERT antibody, is in accordance with other researches which have used the latter antibody (Panarese et al 2006;Mandrioli et al 2007). Those results are now under question since Wu et al (2006) and Zavlaris et al (2009) controvert the specificity of the latter antibody.…”

Section: Resultssupporting

confidence: 83%

“…Recent studies support the notion that the dog is the most appropriate animal model to study human telomerase (Nasir et al 2001;Argyle and Nasir 2003). Canine TERT has been detected immunohistochemically in many canine tumour types using commercially available anti-hTERT antibodies (Renwick et al 2006;Long et al 2006;Panarese et al 2006;Colitz et al 2006;Mandrioli et al 2007;Zarfoss et al 2007;Zavlaris et al 2009) and the evaluation of its expression has now been recognized as an essential approach to the study of cell proliferation and tumorigenesis. Three patterns of immunostaining have been reported to date; nuclear, cytoplasmic and both nuclear and cytoplasmic.…”

Section: Introductionmentioning

confidence: 94%

Immunohistochemical expression of dogTERT in canine testicular tumours in relation to PCNA, ki67 and p53 expression.

et al. 2009

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The objective of the study was to determine the immunohistochemical expression of canine TERT in canine testicular tumours comparing two different antibodies for TERT, and to correlate them with well established markers specific to dividing cells such as PCNA and ki67, and with expression of the p53 tumour suppressor gene. The study included 36 cases of canine testicular tumours, which were categorized as 12 Sertoli Cell Tumours (SCT), 20 seminomas, 3 interstitial cell tumours and 1 mixed germ cell-sex cord stromal tumour (MT). Two antibodies for hTERT were examined; a highly specific TERT antibody, RCK-hTERT, was evaluated for the first time. Immunodetection of RCK-hTERT was observed in 31% of tumours examined (6/20 Seminomas, 4/12 SCT, 1/3 interstitial cell tumour and 0/1 mixed germ cell-sex cord stromal tumour), while the NCL-hTERT in 67% of them (15/20 Seminomas, 6/12 SCT, 3/3 interstitial cell tumour and 0/1 ΜΤ). PCNA immunoreactivity was detected in all cases. Regarding ki67, 3 SCT, 12 seminomas and all interstitial cell tumours showed clear immunoreaction. p53 immunoreactivity was detected in 6 SCT, 15 seminomas and all interstitial cell tumours. The immunohistochemical expression of both TERT antibodies are discussed and compared in order to clarify their potential usefulness in canine testicular malignancies in relation to the expression of well known cell cycle markers. Our results indicate that TERT and PCNA are useful proliferation markers but not helpful to evaluate prognosis. Instead of that ki67 and p53 could be used for predicting aggressiveness in this group of tumours.

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“…Dog liver tissue expressed a single band specific for P-gp and BCRP at the expected molecular weight of 140 kDa and 70 kDa, respectively (see Additional file 3). Anti-p53, -TERT and -ki67 (Mib1) antibodies specificity for dog tissues had been assessed in previous studies [32][33][34][35].…”

Section: Antibodies Validation By Western Blottingmentioning

confidence: 99%

Doxorubicin treatment modulates chemoresistance and affects the cell cycle in two canine mammary tumour cell lines

et al. 2021

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Background Doxorubicin (DOX) is widely used in both human and veterinary oncology although the onset of multidrug resistance (MDR) in neoplastic cells often leads to chemotherapy failure. Better understanding of the cellular mechanisms that circumvent chemotherapy efficacy is paramount. The aim of this study was to investigate the response of two canine mammary tumour cell lines, CIPp from a primary tumour and CIPm, from its lymph node metastasis, to exposure to EC50(20h) DOX at 12, 24 and 48 h of treatment. We assessed the uptake and subcellular distribution of DOX, the expression and function of P-glycoprotein (P-gp) and Breast Cancer Resistance Protein (BCRP), two important MDR mediators. To better understand this phenomenon the effects of DOX on the cell cycle and Ki67 cell proliferation index and the expression of p53 and telomerase reverse transcriptase (TERT) were also evaluated by immunocytochemistry (ICC). Results Both cell lines were able to uptake DOX within the nucleus at 3 h treatment while at 48 h DOX was absent from the intracellular compartment (assessed by fluorescence microscope) in all the surviving cells. CIPm, originated from the metastatic tumour, were more efficient in extruding P-gp substrates. By ICC and qRT-PCR an overall increase in both P-gp and BCRP were observed at 48 h of EC50(20h) DOX treatment in both cell lines and were associated with a striking increase in the percentage of p53 and TERT expressing cells by ICC. The cell proliferation fraction was decreased at 48 h in both cell lines and cell cycle analysis showed a DOX-induced arrest in the S phase for CIPp, while CIPm had an increase in cellular death without arrest. Both cells lines were therefore composed by a fraction of cells sensible to DOX that underwent apoptosis/necrosis. Conclusions DOX administration results in interlinked modifications in the cellular population including a substantial effect on the cell cycle, in particular arrest in the S phase for CIPp and the selection of a subpopulation of neoplastic cells bearing MDR phenotype characterized by P-gp and BCRP expression, TERT activation, p53 accumulation and decrease in the proliferating fraction. Important information is given for understanding the dynamic and mechanisms of the onset of drug resistance in a neoplastic cell population.

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Evaluation of Telomerase in Canine Mammary Tissues by Immunohistochemical Analysis and a Polymerase Chain Reaction-Based Enzyme-Linked Immunosorbent Assay

Cited by 8 publications

References 49 publications

Immunohistochemical expression of h-telomerase reverse transcriptase in canine and feline meningiomas

Immunohistochemical expression of h-telomerase reverse transcriptase in canine and feline meningiomas

Immunohistochemical expression of dogTERT in canine testicular tumours in relation to PCNA, ki67 and p53 expression.

Doxorubicin treatment modulates chemoresistance and affects the cell cycle in two canine mammary tumour cell lines

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