Expression ofRFG/ELE1?/ARA70 in normal and malignant prostatic epithelial cell cultures and lines: Regulation by methylation and sex steroids

Tekur, Seshadri; Lau, Kin‐Mang; Long, J. P.; Burnstein, Kerry L.; Ho, Shuk‐Mei

doi:10.1002/1098-2744(200101)30:1<1::aid-mc1008>3.0.co;2-x

Cited by 27 publications

(24 citation statements)

References 34 publications

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“…AR promoter methylation was excluded as a possible mechanism for the lack of AR expression in primary prostate cell cultures (Grant et al 1996, Tekur et al 2001. Rather, it appears that the androgen-responsive phenotype of human prostatic epithelial cells is dependent on their correct differentiation, which can be maintained via 3D coculture with stroma and/or ECM (see Section 3D in vitro cell culture models to study AR signaling in PCa; Lang et al 2001).…”

Section: Benign Prostate Epithelial Cell Linesmentioning

confidence: 99%

In vitro model systems to study androgen receptor signaling in prostate cancer

Sampson

Neuwirt

Puhr

et al. 2013

Endocrine-Related Cancer

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Prostate cancer (PCa) is one of the most common causes of male cancer-related death in Western nations. The cellular response to androgens is mediated via the androgen receptor (AR), a ligand-inducible transcription factor whose dysregulation plays a key role during PCa development and progression following androgen deprivation therapy, the current mainstay systemic treatment for advanced PCa. Thus, a better understanding of AR signaling and new strategies to abrogate AR activity are essential for improved therapeutic intervention. Consequently, a large number of experimental cell culture models have been established to facilitate in vitro investigations into the role of AR signaling in PCa development and progression. These different model systems mimic distinct stages of this heterogeneous disease and exhibit differences with respect to AR expression/status and androgen responsiveness. Technological advances have facilitated the development of in vitro systems that more closely reflect the physiological setting, for example via the use of three-dimensional coculture to study the interaction of prostate epithelial cells with the stroma, endothelium, immune system and tissue matrix environment. This review provides an overview of the most commonly used in vitro cell models currently available to study AR signaling with particular focus on their use in addressing key questions relating to the development and progression of PCa. It is hoped that the continued development of in vitro models will provide more biologically relevant platforms for mechanistic studies, drug discovery and design ensuring a more rapid transfer of knowledge from the laboratory to the clinic.

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Section: Benign Prostate Epithelial Cell Linesmentioning

confidence: 99%

In vitro model systems to study androgen receptor signaling in prostate cancer

Sampson

Neuwirt

Puhr

et al. 2013

Endocrine-Related Cancer

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“…TSUPr1, a bladder cancer cell line, was maintained as previously described (Tekur et al, 2001). The breast cancer cell line MCF-7 was purchased from ATCC.…”

Section: Cell Culture and 5-azadc Treatmentmentioning

confidence: 99%

“…Amplification of the correct sequences was verified by direct DNA sequencing of PCR product at the Nucleic Acid Facility of University of Massachusetts Medical School. Human 18S ribosomal RNA was amplified from cDNA as a loading control as previously described (Tekur et al, 2001).…”

Section: Rna Isolation and Rt-pcrmentioning

confidence: 99%

“…These genes include RARb2 (Nakayama et al, 2001), ER-a (Lau et al, 2000), pS2 (Lau et al, 2000), androgen receptor (Jarrard et al, 1998;Nakayama et al, 2000), AR-associated-protein 70 (ARA70; Tekur et al, 2001), g-glutamyltransferase (GMT; el Yaagoubi et al, 1998), E-cadherin (E-cad; Graff et al, 1995), P-cad (Jarrard et al, 1997a), RASSF1A (Kuzmin et al, 2002), p16 (Jarrard et al, 1997b), glutathione-S-transferase-Pi (GSTPi; Nelson et al, 1997;Lin et al, 2001;Singal et al, 2001), p14 ARF (Esteller et al, 2000), p27 kip1 (Kibel et al, 2001), progesterone receptor (Lau et al, 2000), caveolin-1 and a-subunit of inhibin (Schmitt et al, 2002), and CD44 (Lou et al, 1999;Verkaik et al, 1999). The biological roles of these genes are diverse; they are all involved in the regulation of basic homeostatic cell mechanisms such as cell cycle regulation, cell-cell interaction, protection against oxidative damage, or tumor suppression.…”

Section: Introductionmentioning

confidence: 99%

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PMP24, a gene identified by MSRF, undergoes DNA hypermethylation-associated gene silencing during cancer progression in an LNCaP model

2004

Oncogene

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Transcriptional silencing of antitumor genes via CpG island methylation could be a mechanism mediating prostate cancer (PCa) progression from an androgensensitive (AS) to an androgen-insensitive (AI) state. We have used the methylation-sensitive restriction fingerprinting (MSRF) technique to identify novel CpG-rich sequences that are differentially methylated between the genome of the AS PCa cell line LNCaP and that of an AI subline LNCaP CS generated by maintaining LNCaP in medium with charcoal-stripped (CS) serum for over 30 passages. One such sequence identified was located on a 5 0 CpG island that was found to span part of the promoter, exon 1, and part of intron 1 of the peroxisomal membrane protein 24 kDa (PMP24) gene. Using semiquantitative RT-PCR and bisulfite genomic sequencing, we established an inverse relationship between mRNA expression and methylation of the 5 0 CpG island of PMP24. PMP24 mRNA was absent in LNCaP CS and the androgen receptor-negative PC-3 cell line; both exhibited dense methylation in the said CpG island. In contrast, PMP24 mRNA was expressed in LNCaP and normal prostatic epithelial cells (NPrECs) whose PMP24 5 0 CpG island remained unmethylated. Treatment of LNCaP CS and PC-3 with the demethylating agent 5-aza-2 0 -deoxycytidine (5-AZAdC) reactivated PMP24 mRNA expression. Transient transfection of PMP24 into LNCaP CS and PC-3 cells induced a significant reduction in cell growth and soft-agar colony formation potential, suggesting that PMP24 gene product has antitumor properties. These results demonstrate the utility of MSRF in the identification of novel, differentially methylated DNA sequences in the genome and suggest that hypermethylation-mediated silencing of PMP24 is an epigenetic event involved in PCa progression to androgen independence.

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“…Inspite of the increasing number of AR coactivators expressed in the prostate, the physiological significance of these cofactors and their role in CaP progression has not been clarified. For example, the AR coactivator ARA70 (Yeh and Chang, 1996) may be involved in prostate carcinogenesis and consists of a key mediator of androgen-oestrogen synergism (Tekur et al, 2001). RAP250, a recently identified nuclear receptor co-activator, is highly expressed in reproductive organs including the prostate and ovary (Caira et al, 2000); however, its role remains unclear.…”

Section: Introductionmentioning

confidence: 99%