Untitled

Moss, Thomas J.

doi:10.1023/a:1006264420822

Cited by 9 publications

(1 citation statement)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although, the molecular mechanisms responsible for regulating mRNA turnover are not completely understood, it is known that many labile mRNAs, including MYCN (22), contain AU-rich elements (ARE) within their 3Ј-untranslated regions (UTRs) (23,24 and reviewed in Ref. 25). At least 14 apparently distinct AREbinding proteins have been identified in eukaryotic cell extracts by UV-cross-linking and gel shift assays (reviewed in Ref.…”

mentioning

confidence: 99%

HuD, a Neuronal-specific RNA-binding Protein, Increases thein Vivo Stability of MYCN RNA

Manohar¹,

Short²,

Nguyen³

et al. 2002

Journal of Biological Chemistry

View full text Add to dashboard Cite

MYCN amplification and consequent deregulated expression plays a crucial role in determining the clinical behavior of neuroblastoma. Enhanced expression of MYCN confers growth potential to neuroblastoma cells, and a direct link between MYCN expression and the development of neuroblastoma has been demonstrated in transgenic mice studies. Although the molecular pathways underlying the regulation of MYCN have not been fully elucidated, post-transcriptional mechanisms appear to be important. Previously, we reported that an embryonic lethal abnormal vision-like (ELAV) protein binds with high specificity to at least two AU-rich elements within the MYCN 3-untranslated region. In this study, we characterized the ability of cis-acting elements within the MYCN 3-untranslated region to destabilize mRNA in cells and examined the functional consequences of its interactions with the ELAV protein HuD. We show that at least 4 cis-acting elements within the MYCN 3-untranslated region are able to signal the degradation of stable heterologous mRNA. Ectopic overexpression of HuD dramatically inhibits RNA decay mediated by the full-length MYCN 3-untranslated region and cis-acting destabilizing elements that harbor HuD binding sites in vivo. HuD may contribute to the malignant phenotype of neuroblastoma cells by stabilizing MYCN mRNA, thereby enhancing steady-state levels of expression of this oncogene. Neuroblastoma (NB),1 a neoplasm that arises from embryonic neural crest tissue, is the second-most common solid pediatric tumor (1). MYCN is amplified in ϳ25% of primary NBs (2-4), and MYCN overexpression is frequently detected in NB tumors (5). Although the clinical significance of MYCN expression in NB tumors that lack MYCN amplification remains controversial (6 -10), a strong correlation between high levels of MYCN expression consequent to genomic amplification of this oncogene and aggressive disease is well established (3-5, 11). Laboratory experiments have further supported an important role for MYCN in determining NB phenotype. Ectopic overexpression of MYCN results in enhanced malignant growth (12, 13), whereas MYCN antisense studies performed by our laboratory and others have shown that down-regulation of MYCN in human NB is associated with a decrease in cellular proliferation and inhibited tumor cell growth in vitro (14 -16). Furthermore, a direct link between MYCN expression and the development of NB has been demonstrated in transgenic mice studies (17).Previously we examined MYCN regulation in N-(neuroblastic, tumorigenic) and S-type (substrate-adherent, non-tumorigenic) subclones (W-N and W-S) of the MYCN-amplified NB cell line NBL-W (18). In addition to distinct morphology and growth characteristics, the subclones have differential levels of MYCN expression despite having the same genomic MYCN copy number. We found that the disparity in steady-state levels of MYCN mRNA in the W-N and W-S cells was largely determined by differences in MYCN mRNA stability (19). Lazarova and co-workers (20) examined MYCN expression in other ...

show abstract

mentioning

confidence: 99%

HuD, a Neuronal-specific RNA-binding Protein, Increases thein Vivo Stability of MYCN RNA

Manohar¹,

Short²,

Nguyen³

et al. 2002

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

Flow cytometry assessment of residual melanoma cells in tumor‐infiltrating lymphocyte cultures

et al. 2012

View full text Add to dashboard Cite

Adoptive transfer of tumor-infiltrating lymphocytes (TIL) is in development for the treatment of metastatic melanoma. In phase II clinical trials, patients with metastatic melanoma that received TIL after preconditioning had a 50-70% clinical response rate. The current approach to generate TIL is to culture melanoma enzyme digests in the presence of IL-2 for a 10-to 20-day period followed by 2 weeks of rapid expansion (REP). Prior to administration, cell therapies are characterized and tested for purity. TIL are characterized by CD3 surface marker expression, and purity is assessed by the amount of tumor remaining in culture. Evaluating TIL purity has traditionally been done by immunohistochemistry, which is often considered semiquantitative. To generate a quantitative assay, we used multiparameter flow cytometry to evaluate the presence of viable tumor cells by staining TIL populations with a viability dye and an antibody cocktail that detects intracellular tumor-antigens gp100, Mart-1, tyrosinase, S100, and surface tumor-antigen melanoma chondroitin sulfate proteoglycan (MCSP), and CD3 on T cells. Tumors were identified by gating on the viable CD3 2 population. Antigens in tumors were initially optimized with individual antibodies using both immunohistochemistry and flow cytometry. When eight different tumor cell lines were spiked into an activated T cell culture, flow cytometry was able to distinguish lymphocytes from tumors in all samples tested. Most importantly, the assay was able to detect melanoma cells in all enzyme digests (9/9) from patient samples. After IL-2-induced TIL expansion, there was a significant decrease in tumor cells; tumor cells were detected in only 2 of 12 samples. In eight IL-2-induced TIL samples that were further expanded in REP, no tumor cells were detected. We have demonstrated that flow cytometry is an alternative to immunohistochemistry for defining the purity of a TIL population. ' 2012 International Society for Advancement of Cytometry

show abstract

Detection of micrometastatic breast cancer by means of real time quantitative RT‐PCR and immunostaining in perioperative blood samples and sentinel nodes

Schröder

Ruiters²,

Jong³

et al. 2003

Intl Journal of Cancer

View full text Add to dashboard Cite

show abstract

Untitled

Cited by 9 publications

References 45 publications

HuD, a Neuronal-specific RNA-binding Protein, Increases thein Vivo Stability of MYCN RNA

HuD, a Neuronal-specific RNA-binding Protein, Increases thein Vivo Stability of MYCN RNA

Flow cytometry assessment of residual melanoma cells in tumor‐infiltrating lymphocyte cultures

Detection of micrometastatic breast cancer by means of real time quantitative RT‐PCR and immunostaining in perioperative blood samples and sentinel nodes

Contact Info

Product

Resources

About