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Vitamin D Receptor Poly(A) Microsatellite Polymorphism and 25-Hydroxyvitamin D Serum Levels: Association with Susceptibility to Breast Cancer.

Colagar AH, Firouzjah HM, Halalkhor S - J Breast Cancer (2015)

Bottom Line: The vitamin D levels of samples were determined by electrochemiluminescence.A larger increase in the risk for breast cancer was found in individuals with the L carrier genotype and lowered 25(OH)D levels.The results primarily suggest that VDR gene polymorphism in the poly(A) microsatellite is associated with 25(OH)D levels and that it can affect the breast cancer risk in the female population from northern Iran.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Cell Biology, University of Mazandaran Faculty of Basic Sciences, Mazandaran, Iran.

ABSTRACT

Purpose: According to previous studies, vitamin D exhibits protective effects against breast cancer via the vitamin D receptor (VDR). There is growing evidence that breast cancer incidence is associated with various polymorphisms of the VDR gene. This study investigates the association of VDR poly(A) microsatellite variants with 25-hydroxyvitamin D (25(OH)D) serum levels and breast cancer risk.

Methods: Polymorphism analysis was performed on a total of 261 blood samples, which were collected from 134 women with breast cancer and 127 controls. Single strand conformation polymorphism was assessed by polymerase chain reaction in combination with sequencing to detect poly(A) lengths for each sample. The vitamin D levels of samples were determined by electrochemiluminescence.

Results: The poly(A) variant L allele frequency was significantly higher in cancer patients than in controls (odds ratio [OR], 1.73; 95% confidence interval [CI], 1.16-2.57; p=0.006). Thus, carriers of the L allele (LS and LL genotypes) have a higher risk for breast cancer (OR, 1.86; 95% CI, 1.13-3.05; p=0.013). A larger increase in the risk for breast cancer was found in individuals with the L carrier genotype and lowered 25(OH)D levels.

Conclusion: The results primarily suggest that VDR gene polymorphism in the poly(A) microsatellite is associated with 25(OH)D levels and that it can affect the breast cancer risk in the female population from northern Iran.

No MeSH data available.


Related in: MedlinePlus

Polymerase chain reaction product in nondenaturing polyacrylamide gel electrophoresis, visualized by silver nitrate staining: SS genotype with ~236-238 bp and LL genotype with ~241-243 bp.
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Figure 2: Polymerase chain reaction product in nondenaturing polyacrylamide gel electrophoresis, visualized by silver nitrate staining: SS genotype with ~236-238 bp and LL genotype with ~241-243 bp.

Mentions: Poly(A) microsatellite genotyping was performed by single-strand conformation polymorphism (SSCP) polymerase chain reaction (PCR). For this purpose the entire genomic sequence of human VDR gene was deduced from the National Center for Biotechnology Information (NCBI) (AC number: NG_008731). Forward and reverse primers were designed by GeneRunner software (Hastings Software, New York, USA). Amplification of DNA fragments was performed by PCR in a final volume of 25 µL containing 1X PCR buffer, 0.8 mM MgCl2, 0.2 mM dNTPs, 0.2 mM of each primer (forward: 5'-CAGTTTGGGAGGTCGAGGTA, and reverse: 5'-TTGTTGTCCAGGTTGGAGAGTAACGG), 30 ng genomic DNA, and 1.25 units Taq DNA polymerase (all materials for PCR purchased from Fermentas, St. Leon-Rot, Germany). PCR was performed according to the following program: initial denaturation for 5 minutes at 94℃ followed by 30 repetitive cycles of denaturation at 94℃ for 30 seconds, annealing at 63.5℃ for 30 seconds and extension at 72℃ for 30 seconds. Final extension temperature was 72℃ for 5 minutes. The PCR products were used for SSCP purposes. Amplification success was detected by 8% polyacrylamide gel visualized by silver nitrate (AgNO3) staining (Figure 2).


Vitamin D Receptor Poly(A) Microsatellite Polymorphism and 25-Hydroxyvitamin D Serum Levels: Association with Susceptibility to Breast Cancer.

Colagar AH, Firouzjah HM, Halalkhor S - J Breast Cancer (2015)

Polymerase chain reaction product in nondenaturing polyacrylamide gel electrophoresis, visualized by silver nitrate staining: SS genotype with ~236-238 bp and LL genotype with ~241-243 bp.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4490260&req=5

Figure 2: Polymerase chain reaction product in nondenaturing polyacrylamide gel electrophoresis, visualized by silver nitrate staining: SS genotype with ~236-238 bp and LL genotype with ~241-243 bp.
Mentions: Poly(A) microsatellite genotyping was performed by single-strand conformation polymorphism (SSCP) polymerase chain reaction (PCR). For this purpose the entire genomic sequence of human VDR gene was deduced from the National Center for Biotechnology Information (NCBI) (AC number: NG_008731). Forward and reverse primers were designed by GeneRunner software (Hastings Software, New York, USA). Amplification of DNA fragments was performed by PCR in a final volume of 25 µL containing 1X PCR buffer, 0.8 mM MgCl2, 0.2 mM dNTPs, 0.2 mM of each primer (forward: 5'-CAGTTTGGGAGGTCGAGGTA, and reverse: 5'-TTGTTGTCCAGGTTGGAGAGTAACGG), 30 ng genomic DNA, and 1.25 units Taq DNA polymerase (all materials for PCR purchased from Fermentas, St. Leon-Rot, Germany). PCR was performed according to the following program: initial denaturation for 5 minutes at 94℃ followed by 30 repetitive cycles of denaturation at 94℃ for 30 seconds, annealing at 63.5℃ for 30 seconds and extension at 72℃ for 30 seconds. Final extension temperature was 72℃ for 5 minutes. The PCR products were used for SSCP purposes. Amplification success was detected by 8% polyacrylamide gel visualized by silver nitrate (AgNO3) staining (Figure 2).

Bottom Line: The vitamin D levels of samples were determined by electrochemiluminescence.A larger increase in the risk for breast cancer was found in individuals with the L carrier genotype and lowered 25(OH)D levels.The results primarily suggest that VDR gene polymorphism in the poly(A) microsatellite is associated with 25(OH)D levels and that it can affect the breast cancer risk in the female population from northern Iran.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Cell Biology, University of Mazandaran Faculty of Basic Sciences, Mazandaran, Iran.

ABSTRACT

Purpose: According to previous studies, vitamin D exhibits protective effects against breast cancer via the vitamin D receptor (VDR). There is growing evidence that breast cancer incidence is associated with various polymorphisms of the VDR gene. This study investigates the association of VDR poly(A) microsatellite variants with 25-hydroxyvitamin D (25(OH)D) serum levels and breast cancer risk.

Methods: Polymorphism analysis was performed on a total of 261 blood samples, which were collected from 134 women with breast cancer and 127 controls. Single strand conformation polymorphism was assessed by polymerase chain reaction in combination with sequencing to detect poly(A) lengths for each sample. The vitamin D levels of samples were determined by electrochemiluminescence.

Results: The poly(A) variant L allele frequency was significantly higher in cancer patients than in controls (odds ratio [OR], 1.73; 95% confidence interval [CI], 1.16-2.57; p=0.006). Thus, carriers of the L allele (LS and LL genotypes) have a higher risk for breast cancer (OR, 1.86; 95% CI, 1.13-3.05; p=0.013). A larger increase in the risk for breast cancer was found in individuals with the L carrier genotype and lowered 25(OH)D levels.

Conclusion: The results primarily suggest that VDR gene polymorphism in the poly(A) microsatellite is associated with 25(OH)D levels and that it can affect the breast cancer risk in the female population from northern Iran.

No MeSH data available.


Related in: MedlinePlus