[Detection of MYCN mRNA in neuroblastoma cell lines by quantitative RT-PCR].
OBJECTIVE
To examine the feasibility and practicability of quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) with SYBR GREEN I fluorescence for detecting the MYCN mRNA expression in neuroblastoma cell line LA-N-5.


METHODS
MYCN mRNA expression in LA-N-5 cells was measured using real time RT-PCR with SYBR GREEN I. Glyceraldehyde phosphate dehydrogenase (GAPDH) was used as internal control. The level of the MYCN mRNA was calculated as MYCN copies/GAPDH copies.


RESULTS
Standard curves were linear and showed high correlations (R2>0.99). The ratio of MYCN mRNA copies to GAPDH mRNA copies was calculated based on specific PCR products. The MYCN mRNA level in LA-N-5 cells was obtained (17.4 +/- 1.2).


CONCLUSIONS
Quantitative RT-PCR with SYBR GREEN I fluorescence may be a sensitive and reliable method for detecting the MYCN mRNA expression. It may also be potential applicable for detecting the MYCN mRNA expression in the small amount neuroblastoma tissues.
Feng
Chen
C
Department of Pediatrics, People's Liberation Army General Hospital, Beijing 100853, China.
Tang
Suo-Qin
SQ
Wang
Jian-Wen
JW
Liu
Li-Zhen
LZ
Gao
Xiao-Ning
XN
Long
Hui
H
D045744:Cell Line, Tumor; D006801:Humans; D009447:Neuroblastoma / Q000378:metabolism* / Q000473:pathology; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D016271:Proto-Oncogene Proteins c-myc; D012333:RNA, Messenger