Sensitive detection of rare Ewing's sarcoma cells in peripheral blood by reverse transcriptase polymerase chain reaction.

Abstract

Disseminated disease is very important in the clinical assessment of pediatric sarcomas. Several reports suggest that reverse transcriptase polymerase chain reaction (RT-PCR) holds great promise in the early staging of cancer patients in general. However, the complexities of these protocols hamper adequate standardization, and their application as routine diagnostic tools has been difficult. The aim of this study is to assess the actual minimal number of tumor cells that may be detected by RT-PCR in a blood sample. Specific tumor cell dilutions from a Ewing's sarcoma cell line reconstituted in peripheral blood from healthy individuals were "ficolled" and submitted to RNA extraction for cDNA preparation and PCR amplification of the t(11-22) (q24;q12) fusion transcript. After PCR amplification, we were able to detect the EWS/FI-1 chimeric gene product at a dilution of 10 tumor cells per 1 or 2 mL of blood. Our simple method supports a role for routine clinical use of RT-PCR in the detection of circulating Ewing's sarcoma cells.

• Authors: M N Fidelia-Lambert, Z Zhuang, M Tsokos
• Journal: Human pathology (Hum Pathol) Vol. 30 Issue 1 Pg. 78-80 (Jan 1999) ISSN: 0046-8177 [Print] United States
• PMID: 9923931 (Publication Type: Journal Article)

Chemical References

• DNA Primers
• DNA, Neoplasm
• EWS-FLI fusion protein
• Oncogene Proteins, Fusion
• Proto-Oncogene Protein c-fli-1
• RNA, Neoplasm
• RNA-Binding Protein EWS
• Transcription Factors

Topics

• Artificial Gene Fusion
• Blotting, Southern
• Bone Neoplasms (diagnosis, genetics)
• DNA Primers (chemistry)
• DNA, Neoplasm (analysis)
• Evaluation Studies as Topic
• Humans
• Neoplastic Cells, Circulating (pathology)
• Oncogene Proteins, Fusion (genetics)
• Proto-Oncogene Protein c-fli-1
• RNA, Neoplasm (isolation & purification)
• RNA-Binding Protein EWS
• Reverse Transcriptase Polymerase Chain Reaction (methods)
• Sarcoma, Ewing (diagnosis, genetics)
• Sensitivity and Specificity
• Transcription Factors (genetics)
• Tumor Cells, Cultured