ABSTRACT Background and Aim: Ewing family of tumors (EFT) encompass a group of small blue round cell tumors, including Ewing sarcoma (ES) and EWSR1- negative undifferentiated small round cell sarcoma. The distinction between the EFTs is essential from a clinical perspective due to prognostic and therapeutic differences and is substantiated by the advent of molecular testing in the modern era. In this study, we tried to characterize EFTs by using fluorescent in situ hybridization (FISH) and reverse transcriptase polymerase chain reaction (RT-PCR). Materials and Methods: A retrospective analysis of 47 cases diagnosed as Ewing sarcoma and undifferentiated small blue round cell tumors over a period of 3 years was done. Immunohistochemistry for CD99, NKX2.2, CCNB3, and DUX4 was performed. EWSR1 and FUS gene rearrangement were performed with locus-specific break apart probes. Further characterization by RT-PCR analysis was attempted by using fusion primers. Results: The mean age of presentation was 21.7 years. 34/47 cases were of skeletal origin, while 13/47 were of extraskeletal origin. Classical morphology was noted in 64% of the cases. All cases were CD99 positive, with variable patterns of expression. DUX4 and CCNB3 IHC were negative in all cases. FISH studies revealed EWSR1 rearrangement in 45/47 cases, while the remaining 2/47 showed FUS rearrangement. Conclusion: In EFTs, EWSR1 gene rearrangement is most common, followed by FUS gene rearrangement. CD99 immunohistochemistry with additional molecular analysis is essential for further classification of EFTs. A detailed diagnostic algorithm including histology, IHC, and molecular studies is imperative for accurate subcategorization of EFTs.
Sable et al. (2026) studied this question.