Cloning, expression, and mutation analysis of NOR1, a novel human gene down-regulated in HNE1 nasopharyngeal carcinoma cell line.
OBJECTIVE
To investigate cloning, expression, and mutation analysis of the putative candidate tumor suppressor gene related with nasopharyngeal carcinoma (NPC).


METHODS
We studied the expression profiles in the NPC cell line HNE(1) with the normal nasopharyngeal epithelial cell as control by using cDNA array representing 11,000 cDNA clusters. EST W95442 was found down-regulated in HNE(1). Subsequently, the corresponding gene sequence including this EST was established by cDNA cloning and the RACE (rapid amplification of cDNA end) procedure. The expression pattern of this gene was examined by using Northern blot analysis in various human tissues. Furthermore, we screened the mutations of the coding sequence of the gene using reverse transcription-polymerase chain reaction and single-strand conformation polymorphisms (RT-PCR-SSCP) as well as direct sequencing analysis.


RESULTS
A novel gene (GenBank accession No. AF462348) was cloned and named NOR(1) standing for oxidored-nitro domain-containing protein 1 (Human Gene Nomenclature Committee-approved symbol). Northern blot analysis revealed that the NOR(1) gene had two transcripts (1.2 kb, 1.6 kb), and expressed ubiquitously in human tissues. Moreover, a Glu58Gly mutation in the exon 1 of NOR(1) was detected in two of 25 NPC biopsies.


CONCLUSIONS
We cloned a novel gene NOR(1), and the Glu58Gly polymorphism of NOR(1) may be involved in the development and/or progression of NPC suggesting that NOR(1) could be a candidate tumor repressor gene related with NPC.
Nie
Xinmin
X
Cancer Research Institute of Xiang-Ya School of Medicine, Central South University, 88 Xiangya Road, 410078, Changsha, Hunan, P.R. China.
Zhang
Bicheng
B
Li
Xiaoling
X
Xiang
Juanjuan
J
Xiao
Bingyi
B
Ma
Jian
J
Zhou
Ming
M
Zhu
Shiguo
S
Lu
Hongbin
H
Gui
Rong
R
Shen
Shourong
S
Li
Guiyuan
G
D000595:Amino Acid Sequence; D001483:Base Sequence; D002899:Chromosomes, Human, Pair 9; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D015536:Down-Regulation; D015870:Gene Expression; D006801:Humans; D026901:Membrane Transport Proteins; D009154:Mutation; D009303:Nasopharyngeal Neoplasms / Q000235:genetics*; D011506:Proteins / Q000235:genetics*; D014407:Tumor Cells, Cultured
D026901:Membrane Transport Proteins; C479458:OSCP1 protein, human; D011506:Proteins