Promoter polymorphisms and transcript levels of nicotinic receptor CHRNA5.
Chromosomal locus 15q25, implicated in lung cancer risk and nicotine dependence, shows extensive linkage disequilibrium that complicates identification of causal variation. Cholinergic receptor nicotinic alpha5 (CHRNA5) has been identified as a lung cancer risk factor. We identified by sequence analysis three haplotypes (delTTC, insATC, and insTGG) in the 5' promoter region and three at the 3'-untranslated region of CHRNA5. Linkage disequilibrium analysis of the 5' variants showed that the insTGG haplotype is associated with three tightly linked risk alleles (nicotine dependence, lung cancer, and chronic obstructive pulmonary disease). The three CHRNA5 promoter haplotypes were statistically significantly associated with lung CHRNA5 transcript levels, determined by real-time polymerase chain reaction. In nontumor lung parenchyma from 68 patients who underwent lung lobectomy, the delTTC haplotype was associated with the highest CHRNA5 transcript levels (relative quantification units = 1.82), whereas the insTGG haplotype was associated with the lowest (0.88 units, P(diff) < .001, Welch t test; all statistical tests were two-sided). Luciferase reporter assays in human lung cancer cell lines A549, H460, H520, and H596 also showed that the 5' region haplotypes were statistically significantly associated with changes in CHRNA5 promoter activity, whereas the 3'-untranslated region variants were not.
Falvella
Felicia S
FS
Department of Predictive and Preventive Medicine, Fondazione IRCCS Istituto Nazionale Tumori, Via G. Venezian 1, Milan, Italy.
Galvan
Antonella
A
Colombo
Francesca
F
Frullanti
Elisa
E
Pastorino
Ugo
U
Dragani
Tommaso A
TA
D002884:Chromosomes, Human, Pair 15; D006239:Haplotypes; D006801:Humans; D015810:Linkage Disequilibrium; D008168:Lung / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011978:Receptors, Nicotinic / Q000235:genetics*; D012307:Risk Factors; D014158:Transcription, Genetic
C447158:CHRNA5 protein, human; D009419:Nerve Tissue Proteins; D011978:Receptors, Nicotinic