Reverse Transcriptase–Polymerase Chain Reaction as an Ancillary Molecular Technique in the Diagnosis of Small Blue Round Cell Tumors by Fine-Needle Aspiration Cytology

Gautam, Upasana; Srinivasan, Radhika; Rajwanshi, Arvind; Bansal, Deepak; Marwaha, Ram Kumar; Vasishtha, Rakesh Kumar

doi:10.1309/ajcppjj0py4xzoec

Cited by 21 publications

(23 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…However, RT-PCR is not available in all institutions and this option is better reserved for difficult cases that could not be resolved by ICC, e.g. Ewing's sarcoma and rhabdomyosarcoma (1 case in our study) [21]. …”

Section: Discussionmentioning

confidence: 99%

Cytologic and Immunophenotypic Features of Malignant Cells in Pediatric Body Fluids

et al. 2015

View full text Add to dashboard Cite

Objective: Cytospin preparations and immunocytochemistry are common methods in hospitals to evaluate malignancies in body fluids. Characteristics of malignant cells in pediatric body fluids have not been adequately evaluated. Study Design: 183 pleural, peritoneal and pericardial pediatric fluid specimens were examined by cytospin preparations and immunocytochemistry from two hospitals using similar procedural techniques. Cytologic diagnoses were correlated with the results of clinical history, histology and ancillary studies. Results: Forty cases with malignancy were identified (21.9%); the most common diagnoses were rhabdomyosarcoma and acute lymphoblastic lymphoma (9 and 8 cases, respectively). Small round cell tumors revealed similar morphology as clusters of small round cells with central nuclei and scant cytoplasm with frequent small vacuoles. Twenty-one cases were evaluated by immunocytochemistry, 12 by flow cytometry and 5 by cytogenetic analysis. CD3, CD20, TdT, CD10, desmin and myogenin were the most common markers. Staining artifacts causing interpretation difficulties were noted in 5 cases that were resolved by molecular studies and deferral for surgical specimens. Conclusions: Small round cell tumors are the most common malignancies encountered in pediatric body fluids and share a nonspecific morphology. Although immunocytochemistry is helpful to arrive at the correct diagnosis, other ancillary studies may be necessary, particularly in hematologic malignancies and other difficult cases.

show abstract

Section: Discussionmentioning

confidence: 99%

Cytologic and Immunophenotypic Features of Malignant Cells in Pediatric Body Fluids

et al. 2015

View full text Add to dashboard Cite

show abstract

“…The most numerous are those associated with the detection of EWRS/FLI [23, 81–87] (Fig. 6) For ERG and ATF1, translocation partners have been also described several primers for its detection through specific RT-PCR [88–91], CREB1 [65, 66], NFTcA2 [77], WT1 [92, 93], ETV4 [94, 95] and CHOP [72, 96–98]. …”

Section: Molecular Techniques For Ewsr1 Chimeric Transcripts Detectionmentioning

confidence: 99%

Molecular detection and targeting of EWSR1 fusion transcripts in soft tissue tumors

et al. 2013

View full text Add to dashboard Cite

Soft tissue tumors are a heterogeneous group of tumors, traditionally classified according to morphology and histogenesis. Molecular classification divides sarcomas into two main categories: (a) sarcomas with specific genetic alterations and (b) sarcomas showing multiple complex karyotypic abnormalities without any specific pattern. Most chromosomal alterations are represented by translocations which are increasingly detected. The identification of fusion transcripts, in fact, not only support the diagnosis but also provides the basis for the development of new therapeutic strategies aimed at blocking aberrant activity of the chimeric proteins. One of the genes most susceptible to breakage/translocation in soft tissue tumors is represented by Ewing sarcoma breakpoint region 1 (EWSR1). This gene has a large number of fusion partners, mainly associated with the pathogenesis of Ewing’s sarcoma but with other soft tissue tumors too. In this review, we illustrate the characteristics of this gene/protein, both in normal cellular physiology and in carcinogenesis. We describe the different fusion partners of EWSR1, the molecular pathways in which is involved and the main molecular biology techniques for the identification of fusion transcripts and for their inhibition.

show abstract

“…Our results are in accord with those of previous studies. 22,32 Nonetheless, molecular techniques have their limits, as evidenced by the 15% (34/222) of cases with uninformative molecular results. Some samples were performed at different institutions, which likely have different methods of sample processing.…”

Section: Discussionmentioning

confidence: 99%

Molecular Diagnosis in Ewing Family Tumors

Gamberi

Cocchi

Benini

et al. 2011

The Journal of Molecular Diagnostics

View full text Add to dashboard Cite

The Ewing's family of tumors (EFTs) are characterized by chimeric transcripts generated by specific chromosomal rearrangements. The most common fusions are between the EWSR1 gene on chromosome 22 and the ETS family of transcription factors; rarely, FUS (on chromosome 16) substitutes for EWSR1. The detection of specific translocations using molecular analysis is now a routine part of the pathological examination of EFT. Here, we report our experience with molecular diagnosis of EFT during the 4 years (2006 -2009) at the Rizzoli Institute. We analyzed 222 consecutive tumors with a presumptive diagnosis of EFT using molecular techniques and IHC. We found five distinct types of EWSR1-FLI1 fusion transcripts resulting from translocation t(11;22), three types of EWSR1-ERG transcripts resulting from t(21;22), and one type of t(2;22) resulting in EWSR1-FEV fusion. Molecular investigation validated 92% of cases ultimately diagnosed as EFT; IHC validated 76% of the cases. Thus, despite the difficulties and limitations associated with both molecular and IHC analysis on fresh and formalin-fixed, paraffin-embedded tissue, a combination of these techniques is the best approach to enhancing the accuracy of EFT diagnosis. We also present our method for choosing which molecular techniques to apply. Finally, we collected the most prevalent breakpoints reported in the literature, indicating which exons are involved, the sequence breakpoints, and the NCBI reference sequences. Ewing's sarcoma (ES) is a highly malignant small round cell tumor (SRCT) that can arise in both bone and soft tissues of children and young adults. Rarely, it occurs also in adults.1,2 It is the second most common pediatric bone tumor after osteosarcoma, accounting for ϳ30% of all primary bone tumors in this age group. 1,3,4 Based on their shared immunophenotypes and molecular parameters, several diagnostic entities that were previously considered distinct are now assembled as a single entity, the Ewing's sarcoma family of tumors (EFTs). These include bone and extraskeletal Ewing's sarcoma, primitive neuroectodermal tumor (PNET), and Askin tumor of the thoracopulmonary region. 1,5-7The histological evaluation of EFT is based on morphological, immunophenotypic, and molecular features. The morphology of ES is variable. These tumors are frequently composed of uniform small round cells with round nuclei containing fine chromatin, scanty clear or eosinophilic cytoplasm, and indistinct cytoplasmic membranes. In less differentiated cases, tumor cells are larger, with prominent nucleoli and irregular contours. The cytoplasm frequently contains PAS-positive diastase-resistant glycogen. PNET is characterized by HomerWright rosettes, which are typical of neuroectodermic differentiation. 6,8,9 Immunohistochemical analysis (IHC), the first ancillary technique used in pathology to confirm the diagnosis of EFT, can distinguish among different variants. The immunohistochemical features of ES are positivity for CD99, FLI1, and caveolin 1 antigens, whereas PNET also shows immun...

show abstract

Reverse Transcriptase–Polymerase Chain Reaction as an Ancillary Molecular Technique in the Diagnosis of Small Blue Round Cell Tumors by Fine-Needle Aspiration Cytology

Cited by 21 publications

References 42 publications

Cytologic and Immunophenotypic Features of Malignant Cells in Pediatric Body Fluids

Cytologic and Immunophenotypic Features of Malignant Cells in Pediatric Body Fluids

Molecular detection and targeting of EWSR1 fusion transcripts in soft tissue tumors

Molecular Diagnosis in Ewing Family Tumors

Contact Info

Product

Resources

About