Korean J Pediatr Hematol Oncol.
2001 Apr;8(1):42-50.
Detection of N-myc Amplification with Differential PCR in Neuroblastoma and It's Clinical Significance
- Affiliations
-
- 1Department of Pediatrics, Yonsei University Wonju College of Medicine, Korea.
- 2Department of Clinical Pathology, Kon Kuk Medical School, Department of Pediatrics, Korea.
- 3Department of Clinical Pathology, Yonsei University College of Medicine, Korea.
- 4Department of Clinical Pathology, Aju University College of Medicine, Korea.
- 5Department of Clinical Pathology, Hallym University College of Medicine,Korea.
- 6Department of Clinical Pathology, Korea University College of Medicine, Korea.
- 7Department of Clinical Pathology, Pusan National University College of Medicine, Korea.
- 8Department of Clinical Pathology, Keimyung University School of Medicine, Korea.
- 9Department of Clinical Pathology, Fatima Hospital, Korea.
- 10Department of General Surgery, Yonsei University College of Medicine, Korea.
- 11Department of General Surgery, Catholic University Medical College, Korea.
- 12Department of Pathology, Yonsei University College of Medicine, Korea.
Abstract
- PURPOSE
The N-myc amplification is one of well known poor prognostic markers in neurblastoma. Because the traditional detection method, Southern blot, is expensive, labor-intensive and time-consuming, the detection of N-myc amplification is not routinely performed in Korea. The purposes of this study are to develop polymerase chain reaction (PCR) for detecting N-myc amplification in neuroblastoma tumor tissue, and to elucidate the clinical significance of N-myc amplification.
METHODS
The clinical data and paraffin embedded tumor specimen of 54 neuroblastoma cases were collected from 10 medical centers in Korea. We have developed semiquantitative method of estimating gene copy number that uses differential PCR. N-myc gene primers (RC N-myc, N-myc 7-1) are amplified together with primers from a single-copy internal control gene (beta-globin). After ethidium bromide-stained agarose gel electrophoresis, the ratio of the two PCR products, which stands for N-myc amplification, is determined. Kaplan-Meier survival analysis was performed to evaluate the prognostic significance of N-myc amplification.
RESULTS
The differential PCR was very effective, less expensive, less labor-intensive, and simple detection method for N-myc amplification. The percentage of N-myc amplification was higher in the patients older than 1 year old (34.1%: 14/41), when they were compared to the patients younger than 1 year old (16.7%: 2/12). The percentage of N-myc amplification was higher in the patients who have primary tumor at adrenal gland (40.9%: 9/22) than who have primary tumor at retroperitoneum (17.6%: 3/17) or at mediastinum (16.7%: 2/12). In Stage I, II, and III patients, the mean survival time of N-myc amplified group was 18 months and that of N-myc umamplified group was 64 months (Log Rank 4.35, P=0.037).
CONCLUSION
The differential PCR was very effective, less expensive, less labor-intensive, and simple detection method for N-myc amplification. The N-myc amplification is one of poor prognostic indicators in Neuroblastoma.
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