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Lack of Association between the Serotonin Transporter (5-HTT) and Serotonin Receptor (5-HT2A) Gene Polymorphisms with Smoking Behavior among Malaysian Malays.

Rozak NI, Ahmad I, Gan SH, Abu Bakar R - Sci Pharm (2014)

Bottom Line: No significant differences in the distribution frequencies of the alleles were found between the smokers and the non-smokers for the 5-HTTLPR polymorphism (x(2) = 0.72, P>0.05) or the 5HT2A polymorphism (x(2) = 0.73, P>0.05).This is the first study conducted on Malaysian Malay males regarding the association of 5-HTTLPR and 5HT2A polymorphisms and smoking behavior.However, the genes were not found to be associated with smoking behavior in our population.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, School of Medical Sciences, Universiti Sains Malaysia, Kelantan, Malaysia.

ABSTRACT
An insertion/deletion polymorphism in the promoter region of the serotonin transporter gene (5-HTTLPR) and a polymorphism (rs6313) in the serotonin 2A receptor gene (5-HT2A) have previously been linked to smoking behavior. The objective of this study was to determine the possible association of the 5-HTTLPR and 5-HT2A gene polymorphisms with smoking behavior within a population of Malaysian male smokers (n=248) and non-smokers (n=248). The 5-HTTLPR genotypes were determined using the polymerase chain reaction (PCR) and were classified as short (S) alleles or long (L) alleles. The 5HT2A genotypes were determined using PCR-restriction fragment length polymorphisms (PCR-RFLP). No significant differences in the distribution frequencies of the alleles were found between the smokers and the non-smokers for the 5-HTTLPR polymorphism (x(2) = 0.72, P>0.05) or the 5HT2A polymorphism (x(2) = 0.73, P>0.05). This is the first study conducted on Malaysian Malay males regarding the association of 5-HTTLPR and 5HT2A polymorphisms and smoking behavior. However, the genes were not found to be associated with smoking behavior in our population.

No MeSH data available.


PCR products for amplification using 5-HTTLPR primers. Lane 1 and 4 show homozygous long allele (L), lane 2 and 5 show heterozygous short-long allele (S/L), while lane 3 and 6 show homozygous short allele (S)
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Figure 1: PCR products for amplification using 5-HTTLPR primers. Lane 1 and 4 show homozygous long allele (L), lane 2 and 5 show heterozygous short-long allele (S/L), while lane 3 and 6 show homozygous short allele (S)

Mentions: Genotyping of the 5-HTTLPR gene polymorphism was performed by using the polymerase chain reaction (PCR) as previously described by Heils et al. [12] with some slight modifications. The PCR reaction was performed in a final volume of 25 μ;l containing 60 ng of genomic DNA, 200 μ;M dNTP mix, 0.24 μ;M of each primer (forward 5'-GGCGTTGCCGCTCTGAATGC-3’ and reverse 5’-GAGGGACTGAGCTGGACAACCAC-3'), 0.75 mM magnesium chloride (MgCl2), 1x ammonium sulfate buffer [(NH4)¬2SO4] (Fermentas, Vilnius, Lithuania), and 1.25 U Taq polymerase (Fermentas, Vilnius, Lithuania). After an initial incubation at 95°C for 2 min, the PCR products were amplified for 35 cycles of denaturation at 95°C for 30 sec, annealing at 62°C for 30 sec, and extension at 72°C for 1 min. The final extension step lasted 7 min at 72°C. The PCR products were then resolved using a 2% agarose gel and were visualized under UV light. Each gel contained one lane of a 100 bp ladder to identify the 528 bp fragment, designated as the L allele, and the 484 bp fragment, designated as the S allele (Fig. 1).


Lack of Association between the Serotonin Transporter (5-HTT) and Serotonin Receptor (5-HT2A) Gene Polymorphisms with Smoking Behavior among Malaysian Malays.

Rozak NI, Ahmad I, Gan SH, Abu Bakar R - Sci Pharm (2014)

PCR products for amplification using 5-HTTLPR primers. Lane 1 and 4 show homozygous long allele (L), lane 2 and 5 show heterozygous short-long allele (S/L), while lane 3 and 6 show homozygous short allele (S)
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4318206&req=5

Figure 1: PCR products for amplification using 5-HTTLPR primers. Lane 1 and 4 show homozygous long allele (L), lane 2 and 5 show heterozygous short-long allele (S/L), while lane 3 and 6 show homozygous short allele (S)
Mentions: Genotyping of the 5-HTTLPR gene polymorphism was performed by using the polymerase chain reaction (PCR) as previously described by Heils et al. [12] with some slight modifications. The PCR reaction was performed in a final volume of 25 μ;l containing 60 ng of genomic DNA, 200 μ;M dNTP mix, 0.24 μ;M of each primer (forward 5'-GGCGTTGCCGCTCTGAATGC-3’ and reverse 5’-GAGGGACTGAGCTGGACAACCAC-3'), 0.75 mM magnesium chloride (MgCl2), 1x ammonium sulfate buffer [(NH4)¬2SO4] (Fermentas, Vilnius, Lithuania), and 1.25 U Taq polymerase (Fermentas, Vilnius, Lithuania). After an initial incubation at 95°C for 2 min, the PCR products were amplified for 35 cycles of denaturation at 95°C for 30 sec, annealing at 62°C for 30 sec, and extension at 72°C for 1 min. The final extension step lasted 7 min at 72°C. The PCR products were then resolved using a 2% agarose gel and were visualized under UV light. Each gel contained one lane of a 100 bp ladder to identify the 528 bp fragment, designated as the L allele, and the 484 bp fragment, designated as the S allele (Fig. 1).

Bottom Line: No significant differences in the distribution frequencies of the alleles were found between the smokers and the non-smokers for the 5-HTTLPR polymorphism (x(2) = 0.72, P>0.05) or the 5HT2A polymorphism (x(2) = 0.73, P>0.05).This is the first study conducted on Malaysian Malay males regarding the association of 5-HTTLPR and 5HT2A polymorphisms and smoking behavior.However, the genes were not found to be associated with smoking behavior in our population.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, School of Medical Sciences, Universiti Sains Malaysia, Kelantan, Malaysia.

ABSTRACT
An insertion/deletion polymorphism in the promoter region of the serotonin transporter gene (5-HTTLPR) and a polymorphism (rs6313) in the serotonin 2A receptor gene (5-HT2A) have previously been linked to smoking behavior. The objective of this study was to determine the possible association of the 5-HTTLPR and 5-HT2A gene polymorphisms with smoking behavior within a population of Malaysian male smokers (n=248) and non-smokers (n=248). The 5-HTTLPR genotypes were determined using the polymerase chain reaction (PCR) and were classified as short (S) alleles or long (L) alleles. The 5HT2A genotypes were determined using PCR-restriction fragment length polymorphisms (PCR-RFLP). No significant differences in the distribution frequencies of the alleles were found between the smokers and the non-smokers for the 5-HTTLPR polymorphism (x(2) = 0.72, P>0.05) or the 5HT2A polymorphism (x(2) = 0.73, P>0.05). This is the first study conducted on Malaysian Malay males regarding the association of 5-HTTLPR and 5HT2A polymorphisms and smoking behavior. However, the genes were not found to be associated with smoking behavior in our population.

No MeSH data available.