Detection of MYCN mRNA in neuroblastoma cell lines by quantitative RT-PCR.
- Author:
Chen FENG
1
;
Suo-Qin TANG
;
Jian-Wen WANG
;
Li-Zhen LIU
;
Xiao-Ning GAO
;
Hui LONG
Author Information
- Publication Type:Journal Article
- MeSH: Cell Line, Tumor; Humans; Neuroblastoma; metabolism; pathology; Proto-Oncogene Proteins c-myc; genetics; RNA, Messenger; analysis; Reverse Transcriptase Polymerase Chain Reaction; methods; Sensitivity and Specificity
- From: Chinese Journal of Contemporary Pediatrics 2007;9(1):47-50
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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.
METHODSMYCN 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.
RESULTSStandard 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).
CONCLUSIONSQuantitative 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.
