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The effect of 5'untranslated region polymorphism in EGF gene, rs4444903, on colorectal cancer.

Chaleshi V, Haghighi MM, Javadi GR, Fatemi SR, Vahedi M, Zali MR - Gastroenterol Hepatol Bed Bench (2013)

Bottom Line: Mutations were confirmed in 10% of the samples by direct sequencing.However, no significant association between this polymorphism and CRC stage was observed (p=0.626).Our data suggest a SNP rs4444903 may not represent a risk factor in the development and progression of CRC among Iranian population.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.

ABSTRACT

Aim: The purpose of this study was to determine the relationship of rs4444903 (EGF+61A/G) SNP genotype with colorectal cancer and tumor stage in an Iranian population.

Background: Epidermal growth factor (EGF) is one of the important proteins that determine survival of cells. EGF binds to its receptor on the cell surface and then activates some of the cell signaling pathway networks within cells that lead to activation or deactivation of factors which are responsible for growth and apoptosis of cells. In this study we assessed the association in EGF polymorphism rs4444903 with colorectal cancer (CRC) in Iranian population.

Patients and methods: We conducted case-control study to investigate the association of polymorphism rs4444903 in EGF, with colorectal cancer risk in Iranian population. Analyzed Polymorphism of EGF rs4444903 with restriction fragment length polymorphisms (RFLP) among two groups of subjects consisting of including 220 cases with colorectal cancer and 220 healthy individuals as controls. Mutations were confirmed in 10% of the samples by direct sequencing.

Results: The frequencies of AA, AG and GG genotypes among cases with colorectal cancer were 28.2, 46.8, and 25.0 % respectively and in controls genotype frequencies were 23.2, 56.4, and 20.5 %, respectively. Frequency of A allele among case group was 51.6% and for control group was 51.4%. The frequency of G allele in case and control was, respectively 48.4% and 48.6% (OR= 1.009, 95% CI= 0.775-1.315; P= 0.946). The percentage of Stage 0, I, II, III, IV were 5%, 9.35%, 38.84%, 30.21% and 16.54%, respectively, among the cases. However, no significant association between this polymorphism and CRC stage was observed (p=0.626).

Conclusion: Our data suggest a SNP rs4444903 may not represent a risk factor in the development and progression of CRC among Iranian population.

No MeSH data available.


Related in: MedlinePlus

Electrophoresis digested products with AluI restricted enzyme on agarose gel showed different bonds. The marker (M) that used was 100 base pairs
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Figure 0001: Electrophoresis digested products with AluI restricted enzyme on agarose gel showed different bonds. The marker (M) that used was 100 base pairs

Mentions: The samples were extracted using standard salting out method and the quality and quantity of DNA was evaluated by Nanodrop Spectrophotometer(20). Genotyping of rs4444903 polymorphism was performed by restriction fragment length polymorphisms analysis. To amplify DNA segment, the specific primers were used (Table 1) (18). The PCR cycle conditions consisted of an initial denaturation step at 95° C for 5 min followed by 30 cycles of 45 s at 95° C; 40 s at 60° C; 45 s at 72° C; and a final elongation at 72° C for 10 min. After PCR, product was run on in to 1% agarose gel in order to ensure a successful reproduction. The products were digested by AluI enzyme (New England Biolabs). The recognition and cutting point of this enzyme is AG/CT, in which SNP region is show by capital letters. Cutting point of enzyme can be observed by a diagonal line. After enzyme digestion, the allele A in 102+91+34+15 bp length, and allele G in 193+34+15 bp lengths were observed. The length of the genotype bands can be seen in the Table 2 (Figure 1). The digested PCR products were determined on a 3% agarose gel and stained with ethidium bromide for visualization under UV light. To confirm genotyping results, a 10% random sample representing all 3 genotypes was sequenced by automated DNA sequencing, using the ABI genetic analyzer 3130xl (Figure 2).


The effect of 5'untranslated region polymorphism in EGF gene, rs4444903, on colorectal cancer.

Chaleshi V, Haghighi MM, Javadi GR, Fatemi SR, Vahedi M, Zali MR - Gastroenterol Hepatol Bed Bench (2013)

Electrophoresis digested products with AluI restricted enzyme on agarose gel showed different bonds. The marker (M) that used was 100 base pairs
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4017510&req=5

Figure 0001: Electrophoresis digested products with AluI restricted enzyme on agarose gel showed different bonds. The marker (M) that used was 100 base pairs
Mentions: The samples were extracted using standard salting out method and the quality and quantity of DNA was evaluated by Nanodrop Spectrophotometer(20). Genotyping of rs4444903 polymorphism was performed by restriction fragment length polymorphisms analysis. To amplify DNA segment, the specific primers were used (Table 1) (18). The PCR cycle conditions consisted of an initial denaturation step at 95° C for 5 min followed by 30 cycles of 45 s at 95° C; 40 s at 60° C; 45 s at 72° C; and a final elongation at 72° C for 10 min. After PCR, product was run on in to 1% agarose gel in order to ensure a successful reproduction. The products were digested by AluI enzyme (New England Biolabs). The recognition and cutting point of this enzyme is AG/CT, in which SNP region is show by capital letters. Cutting point of enzyme can be observed by a diagonal line. After enzyme digestion, the allele A in 102+91+34+15 bp length, and allele G in 193+34+15 bp lengths were observed. The length of the genotype bands can be seen in the Table 2 (Figure 1). The digested PCR products were determined on a 3% agarose gel and stained with ethidium bromide for visualization under UV light. To confirm genotyping results, a 10% random sample representing all 3 genotypes was sequenced by automated DNA sequencing, using the ABI genetic analyzer 3130xl (Figure 2).

Bottom Line: Mutations were confirmed in 10% of the samples by direct sequencing.However, no significant association between this polymorphism and CRC stage was observed (p=0.626).Our data suggest a SNP rs4444903 may not represent a risk factor in the development and progression of CRC among Iranian population.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.

ABSTRACT

Aim: The purpose of this study was to determine the relationship of rs4444903 (EGF+61A/G) SNP genotype with colorectal cancer and tumor stage in an Iranian population.

Background: Epidermal growth factor (EGF) is one of the important proteins that determine survival of cells. EGF binds to its receptor on the cell surface and then activates some of the cell signaling pathway networks within cells that lead to activation or deactivation of factors which are responsible for growth and apoptosis of cells. In this study we assessed the association in EGF polymorphism rs4444903 with colorectal cancer (CRC) in Iranian population.

Patients and methods: We conducted case-control study to investigate the association of polymorphism rs4444903 in EGF, with colorectal cancer risk in Iranian population. Analyzed Polymorphism of EGF rs4444903 with restriction fragment length polymorphisms (RFLP) among two groups of subjects consisting of including 220 cases with colorectal cancer and 220 healthy individuals as controls. Mutations were confirmed in 10% of the samples by direct sequencing.

Results: The frequencies of AA, AG and GG genotypes among cases with colorectal cancer were 28.2, 46.8, and 25.0 % respectively and in controls genotype frequencies were 23.2, 56.4, and 20.5 %, respectively. Frequency of A allele among case group was 51.6% and for control group was 51.4%. The frequency of G allele in case and control was, respectively 48.4% and 48.6% (OR= 1.009, 95% CI= 0.775-1.315; P= 0.946). The percentage of Stage 0, I, II, III, IV were 5%, 9.35%, 38.84%, 30.21% and 16.54%, respectively, among the cases. However, no significant association between this polymorphism and CRC stage was observed (p=0.626).

Conclusion: Our data suggest a SNP rs4444903 may not represent a risk factor in the development and progression of CRC among Iranian population.

No MeSH data available.


Related in: MedlinePlus