The melanocyte inducing factor MITF is stably expressed in cell lines from human clear cell sarcoma

Li, Kim K.C.; Goodall, Jane; Goding, Colin R.; Sk, Liao; Ch, Wang; Lin, Yann-Sheng; Hiraga, Hiroaki; Nojima, Takayuki; Nagashima, Kazuo; Schaefer, Kaitlin; Lee, Kevin A.W.

doi:10.1038/sj.bjc.6601212

Cited by 31 publications

(26 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Elevation of intracellular cAMP leads to PKA activation and finally results in the transcription of MITF; MITF is essential for the expression of tyrosinase, the rate-limiting enzyme in melanogenesis. Whereas the regulatory subunits, type I, ␣ of PKA and MITF are both dramatically up-regulated in our CCSST cell lines, only a subset of these cells also expresses tyrosinase mRNA and protein (35). This may explain the histological presence of immature melanosomal structures but absence of detectable melanin pigment in most CCSST cases.…”

Section: Discussionmentioning

confidence: 83%

See 1 more Smart Citation

Expression Profiling of t(12;22) Positive Clear Cell Sarcoma of Soft Tissue Cell Lines Reveals Characteristic Up-Regulation of Potential New Marker Genes Including ERBB3

et al. 2004

View full text Add to dashboard Cite

Clear cell sarcoma of soft tissue (CCSST), also known as malignant melanoma of soft parts, represents a rare lesion of the musculoskeletal system usually affecting adolescents and young adults. CCSST is typified by a chromosomal t(12;22)(q13;q12) translocation resulting in a fusion between the Ewing sarcoma gene (EWSR1) and activating transcription factor 1 (ATF1), of which the activity in nontransformed cells is regulated by cyclic AMP. Our aim was to identify critical differentially expressed genes in CCSST tumor cells in comparison with other solid tumors affecting children and young adults to better understand signaling pathways regulating specific features of the development and progression of this tumor entity. We applied Affymetrix Human Genome U95Av2 oligonucleotide microarrays representing ϳ12,000 genes to generate the expression profiles of the CCSST cell lines GG-62, DTC-1, KAO, MST2, MST3, and Su-CC-S1 in comparison with 8 neuroblastoma, 7 Ewing tumor, and 6 osteosarcoma cell lines. Subsequent hierarchical clustering of microarray data clearly separated all four of the tumor types from each other and identified differentially expressed transcripts, which are characteristically up-regulated in CCSST. Statistical analysis revealed a group of 331 probe sets, representing ϳ300 significant (P < 0.001) differentially regulated genes, which clearly discriminated between the CCSST and other tumor samples. Besides genes that were already known to be highly expressed in CCSST, like S100A11 (S100 protein) or MITF (microphthalmia-associated transcription factor), this group shows an obvious portion of genes that are involved in cyclic AMP response or regulation, in pigmentation processes, or in neuronal structure and signaling. Comparison with other expression profile analyses on neuroectodermal childhood tumors confirms the high robustness of this strategy to characterize tumor entities based on their gene expression. We found the avian erythroblastic leukemia viral oncogene homologue 3 (ERBB3) to be one of the most dramatically up-regulated genes in CCSST. Quantitative real-time PCR and Northern blot analysis verified the mRNA abundance and confirmed the absence of the inhibitory transcript variant of this gene. The protein product of the member of the epidermal growth factor receptor family ERBB3 could be shown to be highly present in all of the CCSST cell lines investigated, as well as in 18 of 20 primary tumor biopsies. In conclusion, our data demonstrate new aspects of the phenotype and the biological behavior of CCSST and reveal ERBB3 to be a useful diagnostic marker.

show abstract

Section: Discussionmentioning

confidence: 83%

“…35), the CREM, and the protein kinase, cAMP-dependent, regulatory subunit, type I, ␣ (tissue specific extinguisher 1; PRKAR1A).…”

Section: Discussionmentioning

confidence: 99%

Expression Profiling of t(12;22) Positive Clear Cell Sarcoma of Soft Tissue Cell Lines Reveals Characteristic Up-Regulation of Potential New Marker Genes Including ERBB3

et al. 2004

View full text Add to dashboard Cite

show abstract

“…Thus, the structure of EWS-ATF1 and the predicted structure of EWS-CREB1 indicates that mediating cAMP-inducible transcription through PKA-mediated phosphorylation is not a necessary feature of either fusion protein. Interestingly, whereas the MITF-M promoter is cAMPinducible in the presence of SOX10 in melanoma cells through binding of its cAMP response elements by CREB (15), EWS-ATF1 does not seem to transactivate the MITF-M promoter in CCS cells, at least in exogenous constructs (16). Thus, the expression of the MITF-M transcript in CCS may represent a feature of the precursor cell in which the translocation occurs.…”

Section: Discussionmentioning

confidence: 99%

EWS-CREB1: A Recurrent Variant Fusion in Clear Cell Sarcoma—Association with Gastrointestinal Location and Absence of Melanocytic Differentiation

Antonescu

Nafa²,

Segal³

et al. 2006

Clinical Cancer Research

301

267

View full text Add to dashboard Cite

Purpose: Clear cell sarcoma (CCS) usually arises in the lower extremities of young adults and is typically associated with a t(12;22) translocation resulting in the fusion of EWS (EWSR1) with ATF1, a gene encoding a member of the cyclic AMP^responsive element binding protein (CREB) family of transcription factors. CCS arising in the gastrointestinal tract is rare and its pathologic and molecular features are not well defined. Experimental Design: We report a novel variant fusion of EWS to CREB1, a gene at 2q32 encoding another CREB family member highly related to ATF1, detected in three women with gastrointestinal CCS. All three cases contained an identical EWS-CREB1 fusion transcript that was shown by reverse transcription-PCR. In two of the cases tested, EWS gene rearrangement was also confirmed by fluorescence in situ hybridization and the EWS-CREB1 genomic junction fragments were isolated by long-range DNA PCR. Results: Morphologically, all three tumors lacked melanin pigmentation. By immunohistochemistry, there was a strong and diffuse S100 protein reactivity, whereas all melanocytic markers were negative. Ultrastructurally, two of the cases lacked melanosomes. The melanocyte-specific transcript of MITF was absent in two cases, and only weakly expressed in the third case. The Affymetrix gene expression data available in one case showed lower expression of the melanocytic genes MITF, TYR, and TYRP1, compared with four EWS-ATF1-positive CCSs of non-gastrointestinal origin. Conclusions: EWS-CREB1 may define a novel subset of CCS that occurs preferentially in the gastrointestinal tract and shows little or no melanocytic differentiation. Thus, evidence of melanocytic lineage or differentiation is not a necessary feature of sarcomas with gene fusions involving CREB family members.Clear cell sarcoma (CCS), also known as melanoma of soft parts, typically presents in the deep soft tissues of the lower extremity, in close proximity to tendons, fascia, or aponeuroses. Young adults are preferentially affected and the clinical course is often marked by regional and distant metastases. Most CCS show immunoreactivity for melanoma markers, such as HMB45, and contain melanosomes. Indeed, most CCS share a melanocytic gene expression signature with melanomas (1). However, CCS are also genetically distinct from melanomas, as they lack BRAF mutations (2) and show, in most cases, a recurrent chromosomal translocation t(12;22)(q13;q12), resulting in the fusion of the EWS gene (also known as EWSR1) on 22q12 with the activating transcription factor-1 gene (ATF1) on 12q13 (3 -6).Primary CCSs of the gastrointestinal tract are rare (7,8). Gastrointestinal CCS includes a histologic variant rich in osteoclast-type giant cells which uniformly express S100 protein, but lack melanocytic differentiation by immunohistochemistry, being negative for HMB45 and Mart-1 (9). As a result of its rarity in the gastrointestinal tract, the differential diagnosis of CCS in this site includes more common mesenchymal or neuroectodermal neoplasms,...

show abstract

“…ST-CCS cell lines MST1 and Kao are previously described (Li et al, 2003). For selection of stable transformants DNA was introduced by calcium phosphate/DNA co-precipitation, using 10 μg of pSilencer 2.1-U6neo plasmid as positive control (or test shRNA plasmid) and 10 μg of pGem3 as carrier.…”

Section: Cell Lines and Cell Culturementioning

confidence: 99%

The RNA Pol II sub-complex hsRpb4/7 is required for viability of multiple human cell lines

Zhao¹,

Li²,

Ng³

et al. 2012

Protein Cell

View full text Add to dashboard Cite

The evolutionarily conserved RNA Polymerase II Rpb4/7 sub-complex has been thoroughly studied in yeast and impacts gene expression at multiple levels including transcription, mRNA processing and decay. In addition Rpb4/7 exerts differential effects on gene expression in yeast and Rpb4 is not obligatory for yeast (S. cerevisiae) survival. Specialised roles for human (hs) Rpb4/7 have not been extensively described and we have probed this question by depleting hsRpb4/7 in established human cell lines using RNA interference. We find that Rpb4/7 protein levels are inter-dependent and accordingly, the functional effects of depleting either protein are co-incident. hsRpb4/7 exhibits gene-specific effects and cells initially remain viable upon hsRpb4/7 depletion. However prolonged hsRpb4/7 depletion is cytotoxic in the range of cell lines tested. Protracted cell death occurs by an unknown mechanism and in some cases is accompanied by a pronounced elongated cell morphology. In conclusion we provide evidence for a gene-specific role of hsRpb4/7 in human cell viability.

show abstract

The melanocyte inducing factor MITF is stably expressed in cell lines from human clear cell sarcoma

Cited by 31 publications

References 44 publications

Expression Profiling of t(12;22) Positive Clear Cell Sarcoma of Soft Tissue Cell Lines Reveals Characteristic Up-Regulation of Potential New Marker Genes Including ERBB3

Expression Profiling of t(12;22) Positive Clear Cell Sarcoma of Soft Tissue Cell Lines Reveals Characteristic Up-Regulation of Potential New Marker Genes Including ERBB3

EWS-CREB1: A Recurrent Variant Fusion in Clear Cell Sarcoma—Association with Gastrointestinal Location and Absence of Melanocytic Differentiation

The RNA Pol II sub-complex hsRpb4/7 is required for viability of multiple human cell lines

Contact Info

Product

Resources

About