Characterization of a novel telomerase-immortalized human endometrial stromal cell line, St-T1b

Samalecos, Annemarie; Reimann, Katja; Wittmann, Stefanie; Schulte, Heinrich M.; Brosens, Jan J.; Bamberger, Ana‐Maria; Gellersen, Birgit

doi:10.1186/1477-7827-7-76

Cited by 64 publications

(62 citation statements)

References 38 publications

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“…Various immortalized cell lines were established after lentiviral transduction of plasmids expressing either SV40 large T antigen [19,27,28] or telomerase [20,29,30]. In the presence of cAMP these cell lines were shown to differentiate into a cobblestone-like phenotype and to express marker genes of decidualization such as IGFBP1, PRL and FOXO1A [30]. The cAMP-dependent induction of PRL and IGFBP1 in THESC analysed here confirms these previous observations.…”

Section: Discussionsupporting

confidence: 84%

“…Indeed, upon cAMP treatment MPA-dependent induction of PRL has been observed in HIESC-2 and St-TIb cultures [28,30] and similar to primary HESC [12] the combined stimulation was more efficient than cAMP treatment alone [30]. Moreover, MPA-induced expression of PRL and IGFBP1 in absence of cAMP has also been noticed in immortalized cultures [20,29] suggesting a physiological, PR-dependent response.…”

Section: Discussionmentioning

confidence: 99%

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Evaluation of human first trimester decidual and telomerase-transformed endometrial stromal cells as model systems of in vitro decidualization

Saleh

Otti

Fiala³

et al. 2011

Reprod Biol Endocrinol

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BackgroundDecidualization, the differentiation process of maternal uterine stromal cells into secretory decidual cells, is a prerequisite for successful implantation and progression of pregnancy. For in vitro differentiation mostly primary human endometrial stromal cells (HESC) isolated from uterine samples after hysterectomy for benign gynaecological diseases are utilised. However, a continuous supply of endometrial tissue is often lacking. Hence, we analysed whether cultivated human decidual stromal cells (HDSC) prepared from first trimester pregnancy terminations may represent an alternative model system for in vitro decidualization. Moreover, based on the expression of critical marker genes these cells were compared to a previously established endometrial stromal cell line during in vitro differentiation.MethodsHDSC isolated from decidual tissue attached to first trimester placentae, and telomerase-transformed human endometrial stromal cells (THESC) were characterised by immunofluorescence and differentiated in vitro using either cyclic adenosine monophosphate (cAMP) and/or estrogen (E2)/progesterone (P4). Proliferation was measured by analyzing cumulative cell numbers. Expression of mRNAs encoding progesterone receptor (PR), prolactin (PRL), insulin-like growth factor binding protein-1 (IGFBP1), and Dickkopf-1 (DKK1) was evaluated using quantitative PCR after 3, 6, 9 and 12 days of in vitro differentiation. PRL and IGFBP-1 protein expression was investigated by enzyme-linked immunosorbent assay (ELISA) and Western blotting, respectively. Furthermore, forkhead box O1A (FOXO1A), a critical transcription factor in decidualization, was analysed by immunofluorescence and Western blotting at two different time points of differentiation.ResultsTreatment with cAMP provoked morphological changes and growth arrest of THESC and HDSC, the latter showing loss of cells after 6 days of treatment. E2P4 stimulation did neither affect cell morphology nor proliferation of THESC and HDSC. Upon cAMP stimulation PR mRNA was suppressed in HDSC but not in THESC, whereas E2P4 did not alter transcript levels in both cell types. Protein expression of PR-A and PR-B was detectable in HDSC and diminished under cAMP, whereas THESC failed to produce the nuclear receptors. Supplementation of cAMP induced mRNA and protein expression of PRL and IGFBP-1 in both cell types at day 3, 6, 9, and 12 of treatment. In HDSC stimulation with E2P4 increased PRL and IGFBP-1 mRNA and protein production, whereas hormone treatment did not induce the two factors in THESC. E2P4 increased DKK1 mRNA at all time points in HDSC and cAMP provoked induction at day 9 and 12 of differentiation. In contrast, cAMP suppressed DKK1 mRNA in THESC, whereas E2P4 was ineffective. In both cell types combined treatments with cAMP and E2P4 provoked higher expression levels of PRL and IGFBP1 mRNA and protein as compared to cAMP stimulation alone. FOXO1A protein and its nuclear abundance were increased by cAMP in both cell types. However, reduction of its nuclear localisation upo...

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Section: Discussionsupporting

confidence: 84%

Section: Discussionmentioning

confidence: 99%

Evaluation of human first trimester decidual and telomerase-transformed endometrial stromal cells as model systems of in vitro decidualization

Saleh

Otti

Fiala³

et al. 2011

Reprod Biol Endocrinol

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“…Expression of miR-142-3p was up-and downregulated in the immortalized endometrial stroma cell line St-T1b (Samalecos et al, 2009) and in primary endometrial stroma cells obtained from ectopic endometrial explants of six patients with endometriosis, classified rASRM stadium III or IV (Schneider et al, 2013) (Ethical approval number 1 IX Greb I, dated 6 December 2012). Changes in predicted target gene expression were studied by quantitative polymerase chain reaction (PCR) and Western blotting.…”

Section: Methodsmentioning

confidence: 99%

miR-142-3p is a novel regulator of cell viability and proinflammatory signalling in endometrial stroma cells

Kästingschäfer

Schäfer

Kiesel

et al. 2015

Reproductive BioMedicine Online

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Endometriosis is associated with severe pelvic pain and reduced fertility. Recently, it has been linked to a dysregulation of microRNAs (miRNAs), which are post-transcriptional regulators of gene expression. The functional effect of dysregulated miR-142-3p expression in endometrial stroma cells was investigated. An increased expression of miR-142-3p resulted in a significantly reduced expression of steroid sulfatase and interleukin-6-coreceptor gp130 as well as reduced interleukin-6-mediated activation of the STAT3-pathway, suggesting an effect of miR-142-3p both on steroid hormone- and cytokine-mediated signalling events. At the functional level, miR-142-3p overexpression significantly reduced cell viability (P ≤ 0.01). miR-142-3p regulation emerges as a future therapeutic strategy for endometriosis.

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“…Telomerase activity is absent in the majority of human somatic cells but present in immortalized cell lines, stem and germ cells and in approximately 90% of human cancers 15 . There are studies suggesting that the progression of malignant diseases depends on telomerase reactivation, and that an inhibiting agent of this enzyme could be an effective antitumor drug 7,15 .…”

Section: Introductionmentioning

confidence: 99%

“…There are studies suggesting that the progression of malignant diseases depends on telomerase reactivation, and that an inhibiting agent of this enzyme could be an effective antitumor drug 7,15 .…”

Section: Introductionmentioning

confidence: 99%

AZT on telomerase activity and cell proliferation in HS 839.T melanoma cells

Sobrinho¹,

Gragnani

Santos³

et al. 2012

Acta Cir. Bras.

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PURPOSE:To evaluate telomerase activity and proliferation of HS839.T melanoma cells, subjected to the action of AZT. METHODS:Cells were grown in triplicate, AZT at different concentrations: 50, 100 and 200μM, was added and left for 24 and 48 hours, and its effects were compared with the control group. Telomerase activity was detected by PCR and cell proliferation was evaluated by MTT. RESULTS:After 24 hours, there was no inhibition of cell proliferation or telomerase activity when compared to the control group. After 48 hours, there was a momentary decrease, suggesting that the cell lines used in this study are sensitive to AZT, but quickly recover both the enzyme activity and cell proliferation. CONCLUSION:The action of AZT on the melanoma cells studied, at the concentrations and times tested, did not inhibit telomerase activity nor affect cell proliferation.Key words: Telomerase. Melanoma. Zidovudine. Cell Proliferation. In Vitro.RESUMO OBJETIVO: Avaliar a atividade da telomerase e da proliferação de células de melanoma HS839.T submetidas à ação do AZT. MÉTODOS:As células foram cultivadas, em triplicata, com diferentes concentrações de AZT: 50, 100 e 200µM, por 24h e 48h, seus efeitos comparados com o grupo controle. A atividade da telomerase foi detectada por PCR e a proliferação celular avaliada por MTT. RESULTADOS:No tempo de 24 horas, não houve inibição da proliferação celular e da atividade da telomerase em comparação com o grupo controle. No período de 48 horas, houve uma diminuição momentânea, sugerindo que as células das linhagens utilizadas neste estudo são sensíveis ao AZT, mas que recuperam a atividade enzimática e proliferativa.CONCLUSÃO: Nas células de melanoma HS839.T estudadas e nas concentrações e tempos propostos, a ação do AZT não inibiu a atividade da telomerase e não afetou a proliferação celular.Descritores: Telomerase. Melanoma. Zidovudina. Proliferação de Células. In Vitro.Souza Sobrinho CP et al.

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Characterization of a novel telomerase-immortalized human endometrial stromal cell line, St-T1b

Cited by 64 publications

References 38 publications

Evaluation of human first trimester decidual and telomerase-transformed endometrial stromal cells as model systems of in vitro decidualization

Evaluation of human first trimester decidual and telomerase-transformed endometrial stromal cells as model systems of in vitro decidualization

miR-142-3p is a novel regulator of cell viability and proinflammatory signalling in endometrial stroma cells

AZT on telomerase activity and cell proliferation in HS 839.T melanoma cells

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