Establishment of a new human cell line (EN) with TP53 mutation derived from endometrial carcinoma

Isaka, Keiichi; Nishi, Hirotaka; Sagawa, Yasukazu; Nakada, Toshihide; Osakabe, Yasuhiro; Serizawa, Hiromi; Ebihara, Yoshiro; Takayama, M.

doi:10.1016/s0165-4608(02)00673-8

Cited by 14 publications

(17 citation statements)

References 16 publications

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“…By contrast, established human endometrial carcinoma cell lines frequently employed in studies on EEC physiology have lost normal epithelial anchorage-dependent growth control (Isaka et al 2003). Cellular polarization is a critical parameter affecting numerous cell functions (Yeaman et al 1999) and has been shown in primary EECs to be important for embryo attachment and implantation (Meseguer et al 2001) and to enhance protein secretion (Negami & Tominaga 1989).…”

Section: Discussionmentioning

confidence: 99%

“…The human endometrial carcinoma cell lines Ishikawa, RL-95, ECC-1, KLE, HEC-1A and EN revealed altered or impaired hormonal responsiveness (Thie et al 1995, Jazaeri et al 2001, Dardes et al 2002, Di Nezza et al 2003, Farnell & Ing 2003, Isaka et al 2003. In hTERT-EECs, the level of expression of ER-alpha, but not ER-beta, was regulated by estrogen, demonstrating different regulatory processes to affect the transcriptional activation of the two human ER isoforms in hTERT-EECs.…”

Section: Steroid Hormone-responsive Htert-eecs · S Hombach-klonisch Amentioning

confidence: 99%

“…Endometrial carcinoma cell lines, including ECC-1, HEC-1A, RL-95, Ishikawa and EN, have long been employed as experimental models but their usefulness is limited since their transformed phenotype has partially resulted in a loss of physiological growth regulation and polarization or altered steroid hormone receptor regulation (Thie et al 1995, Koopman et al 1999, Di Nezza et al 2003, Farnell & Ing 2003, Isaka et al 2003. In particular, primary endometrial epithelial cells (EECs) display a rapid decrease in proliferative capacity within a few culture passages (Mulholland et al 1988, ClassenLinke et al 1997, Arnold et al 2001.…”

Section: Introductionmentioning

confidence: 99%

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Regulation of functional steroid receptors and ligand-induced responses in telomerase-immortalized human endometrial epithelial cells

Hombach‐Klonisch¹,

Kehlen²,

Fowler³

et al. 2005

Journal of Molecular Endocrinology

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Information on the regulation of steroid hormone receptors and their distinct functions within the human endometrial epithelium is largely unavailable. We have immortalized human primary endometrial epithelial cells (EECs) isolated from a normal proliferative phase endometrium by stably transfecting the catalytic subunit (hTERT) of the human telomerase complex and cultured these hTERT-EECs now for over 350 population doublings. Active hTERT was detected in hTERT-EECs employing the telomerase repeat amplification assay protocol. hTERT-EECs revealed a polarized, non-invasive epithelial phenotype with apical microvilli and production of a basal lamina when grown on a three-dimensional collagen-fibroblast lattice. Employing atomic force microscopy, living hTERT-EECs were shown to produce extracellular matrix (ECM) components and ECM secretion was modified by estrogen and progesterone (P4). hTERT-EECs expressed inducible and functional endogenous estrogen receptor-alpha (ER-alpha) as demonstrated by estrogen response element reporter assays and induction of P4 receptor (PR). P4 treatment down-regulated PR expression, induced MUC-1 gene activity and resulted in increased ER-beta transcriptional activity. Gene activities of cytokines and their receptors interleukin (IL)-6, leukemia inhibitory factor (LIF), IL-11 and IL-6 receptor (IL6-R), LIF receptor and gp130 relevant to implantation revealed a 17 beta-estradiol (E2)-mediated up-regulation of IL-6 and an E2-and P4-mediated up-regulation of IL6-R in hTERT-EECs. Thus, hTERT-EECs may be regarded as a novel in vitro model to investigate the role of human EECs in steroid hormone-dependent normal physiology and pathologies, including implantation failure, endometriosis and endometrial cancer.

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Section: Discussionmentioning

confidence: 99%

Section: Steroid Hormone-responsive Htert-eecs · S Hombach-klonisch Amentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Regulation of functional steroid receptors and ligand-induced responses in telomerase-immortalized human endometrial epithelial cells

Hombach‐Klonisch¹,

Kehlen²,

Fowler³

et al. 2005

Journal of Molecular Endocrinology

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“…Protein extraction for telomerase assays was performed according to protocols published previously, with minor modifications (16,14). Briefly, cells were harvested in CHAPS (3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate) lysis buffer (Intergen, Gaithersburg, Md.)…”

Section: Samplesmentioning

confidence: 99%

Hypoxia-Inducible Factor 1 Mediates Upregulation of Telomerase (hTERT)

Nishi

Nakada

Kyo

et al. 2004

Molecular and Cellular Biology

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189

136

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Hypoxia occurs during the development of the placenta in the first trimester and correlates with both trophoblast differentiation and the induction of telomerase activity through hTERT expression. We sought to determine the mechanism of regulation of hTERT expression during hypoxia. We show that hypoxia-inducible factor 1␣ (HIF-1␣) and hTERT expression in the human placenta decrease with gestational age and that these are overexpressed in preeclamptic placenta, a major complication of pregnancy. Hypoxia not only transactivates the hTERT promoter activity but also enhances endogenous hTERT expression. The hTERT promoter region between ؊165 and ؉51 contains two HIF-1 consensus motifs, and in vitro reporter assays show that these are essential for hTERT transactivation by HIF-1. Introduction of an antisense oligonucleotide for HIF-1 diminishes hTERT expression during hypoxia, indicating that upregulation of hTERT by hypoxia is directly mediated through HIF-1. Our results provide persuasive evidence that the regulation of hTERT promoter activity by HIF-1 represents a mechanism for trophoblast growth during hypoxia and suggests that this may be a generalized response to hypoxia in various human disorders including resistance to cancer therapeutics by upregulating telomerase.

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“…This is likely due to difficulties in growing sufficient quantities of primary epithelial cells from the endocervix because of its relatively small surface area. Limitations to establishing appropriate in vitro endocervix-derived epithelial cell models are also likely compounded by more generic issues that can arise from the passage of primary and transformed epithelial cell lines derived from FRT tissues, including loss or change in hormone receptor expression, hormone responsiveness, and anti-microbial molecule expression (Isaka et al, 2003; Joly et al, 2009). …”

Section: Introductionmentioning

confidence: 99%

Innate immune mediator profiles and their regulation in a novel polarized immortalized epithelial cell model derived from human endocervix

Buckner

Schust

Ding³

et al. 2011

Journal of Reproductive Immunology

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The endocervix in the female reproductive tract (FRT) is susceptible to sexually transmitted pathogens such as Chlamydia trachomatis and Neisseria gonorrhoeae. Endocervical epithelial cells in vivo make innate immune mediators that likely aid in the protection from these pathogens. In vitro studies to investigate the innate epithelial cell immune response to endocervical pathogens have been hindered by the paucity of human endocervix-derived epithelial cell lines that display the differentiation proteins and functional characteristics of their site of origin. We have established an immortalized epithelial cell line (A2EN) derived from an endocervical tissue explant that can be polarized to exhibit distinct apical and basolateral membrane domains. Polarized A2EN cells secrete mucus at their apical surface, and express MUC5B, a mucin specific to the endocervix. Polarized A2EN cells also express hormone receptors that respond appropriately to female steroid hormones. Polarized A2EN cells can be stimulated with the toll-like receptor 3 agonist, polyI:C, to express anti-microbial peptides (AMPs) as well as pro-inflammatory cytokines and chemokines. Cytokines and chemokines are also differentially secreted depending on the hormone milieu in which the cells are exposed. We conclude that polarized A2EN cells maintain distinctive phenotypic and functional characteristics of the epithelial cells found in the endocervix and, hence, could provide a useful, new in vitro model system for investigations on the role of endogenous and exogenous factors that regulate endocervical epithelial cell immunity including studies on sexually transmitted infections and topical microbicides.

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Establishment of a new human cell line (EN) with TP53 mutation derived from endometrial carcinoma

Cited by 14 publications

References 16 publications

Regulation of functional steroid receptors and ligand-induced responses in telomerase-immortalized human endometrial epithelial cells

Regulation of functional steroid receptors and ligand-induced responses in telomerase-immortalized human endometrial epithelial cells

Hypoxia-Inducible Factor 1 Mediates Upregulation of Telomerase (hTERT)

Innate immune mediator profiles and their regulation in a novel polarized immortalized epithelial cell model derived from human endocervix

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