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The -786T>C promoter polymorphism of the NOS3 gene is associated with prostate cancer progression.

Marangoni K, Araújo TG, Neves AF, Goulart LR - BMC Cancer (2008)

Bottom Line: NOS3 gene expression levels were affected by the -786T>C polymorphism, and there was a 2-fold increase in NOS3 levels favored by the incorporation of each C allele.NOS3 levels higher than 80% of the constitutive gene expression level (B2M) presented a 4-fold increase in PCa occurrence.The -786T>C polymorphism was the most important promoter alteration of the NOS3 gene that may affect the PCa progression, but not its occurrence, and the incorporation of the C allele is associated with increased levels of NOS3 transcripts.

View Article: PubMed Central - HTML - PubMed

Affiliation: Federal University of Uberlândia, Institute of Genetics and Biochemistry, Molecular Genetics Laboratory, Campus Umuarama, Block 2E, Room 24, 38400-902, Uberlândia, MG, Brazil. kmarangoni@yahoo.com.br

ABSTRACT

Background: There is no biological or epidemiological data on the association between NOS3 promoter polymorphisms and prostate cancer. The polymorphisms in the promoter region of NOS3 gene may be responsible for variations in the plasma NO, which may promote cancer progression by providing a selective growth advantage to tumor cells by angiogenic stimulus and by direct DNA damage.

Methods: This study aimed evaluating the NOS3 promoter polymorphisms by PCR-SSCP and sequencing, associating genotypes and haplotypes with NOS3 expression levels through semi-quantitative RT-PCR, and with PCA3 mRNA detection, a specific tumor biomarker, in the peripheral blood of pre-surgical samples from 177 patients; 83 PCa and 94 BPH.

Results: Three novel SNPs were identified -764A>G, -714G>T and -649G>A in the NOS3 gene promoter region, which together with the -786T>C generated four haplotypes (N, T, C, A). NOS3 gene expression levels were affected by the -786T>C polymorphism, and there was a 2-fold increase in NOS3 levels favored by the incorporation of each C allele. NOS3 levels higher than 80% of the constitutive gene expression level (B2M) presented a 4-fold increase in PCa occurrence.

Conclusion: The -786T>C polymorphism was the most important promoter alteration of the NOS3 gene that may affect the PCa progression, but not its occurrence, and the incorporation of the C allele is associated with increased levels of NOS3 transcripts. The NOS3 transcript levels presented a bimodal behavior in tumor development and may be used as a biomarker together with the PCA3 marker for molecular staging of the prostate cancer.

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Related in: MedlinePlus

Nested RT-PCR for the PCA3 and B2M gene expression analysis. A) Lanes 1 to 4: amplifications for the PCA3 gene, where sample pairs 1 and 2, 3 and 4, correspond to two RNA extraction replicates for each patient. The detection of the expected 277-bp PCA3 fragment evidences the presence of circulating tumor cells. Lane M: 100-bp size marker. B) Lanes 1 to 4: amplifications for the B2M gene (534-bp). Semi-quantitative RT-PCR for the NOS3 and B2M gene expression analysis. The 322-bp and 534-bp fragments correspond to the NOS3 and B2M genes, respectively. Lane M: 100-bp size marker. Lanes 1 to 10 represent individual patients, where (C) represent BPH patients' profile, and (D) the PCa patients' profile.
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Figure 3: Nested RT-PCR for the PCA3 and B2M gene expression analysis. A) Lanes 1 to 4: amplifications for the PCA3 gene, where sample pairs 1 and 2, 3 and 4, correspond to two RNA extraction replicates for each patient. The detection of the expected 277-bp PCA3 fragment evidences the presence of circulating tumor cells. Lane M: 100-bp size marker. B) Lanes 1 to 4: amplifications for the B2M gene (534-bp). Semi-quantitative RT-PCR for the NOS3 and B2M gene expression analysis. The 322-bp and 534-bp fragments correspond to the NOS3 and B2M genes, respectively. Lane M: 100-bp size marker. Lanes 1 to 10 represent individual patients, where (C) represent BPH patients' profile, and (D) the PCa patients' profile.

Mentions: A sensitive nested PCR assay for the detection of PCA3 mRNA was performed as previously reported [20] with minor modifications. A total of 137 patients were analyzed for the expression of circulating tumor cells in the peripheral blood, as evidenced by the PCA3 tumor biomarker positivity (Figure 3A and 3B).


The -786T>C promoter polymorphism of the NOS3 gene is associated with prostate cancer progression.

Marangoni K, Araújo TG, Neves AF, Goulart LR - BMC Cancer (2008)

Nested RT-PCR for the PCA3 and B2M gene expression analysis. A) Lanes 1 to 4: amplifications for the PCA3 gene, where sample pairs 1 and 2, 3 and 4, correspond to two RNA extraction replicates for each patient. The detection of the expected 277-bp PCA3 fragment evidences the presence of circulating tumor cells. Lane M: 100-bp size marker. B) Lanes 1 to 4: amplifications for the B2M gene (534-bp). Semi-quantitative RT-PCR for the NOS3 and B2M gene expression analysis. The 322-bp and 534-bp fragments correspond to the NOS3 and B2M genes, respectively. Lane M: 100-bp size marker. Lanes 1 to 10 represent individual patients, where (C) represent BPH patients' profile, and (D) the PCa patients' profile.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2571109&req=5

Figure 3: Nested RT-PCR for the PCA3 and B2M gene expression analysis. A) Lanes 1 to 4: amplifications for the PCA3 gene, where sample pairs 1 and 2, 3 and 4, correspond to two RNA extraction replicates for each patient. The detection of the expected 277-bp PCA3 fragment evidences the presence of circulating tumor cells. Lane M: 100-bp size marker. B) Lanes 1 to 4: amplifications for the B2M gene (534-bp). Semi-quantitative RT-PCR for the NOS3 and B2M gene expression analysis. The 322-bp and 534-bp fragments correspond to the NOS3 and B2M genes, respectively. Lane M: 100-bp size marker. Lanes 1 to 10 represent individual patients, where (C) represent BPH patients' profile, and (D) the PCa patients' profile.
Mentions: A sensitive nested PCR assay for the detection of PCA3 mRNA was performed as previously reported [20] with minor modifications. A total of 137 patients were analyzed for the expression of circulating tumor cells in the peripheral blood, as evidenced by the PCA3 tumor biomarker positivity (Figure 3A and 3B).

Bottom Line: NOS3 gene expression levels were affected by the -786T>C polymorphism, and there was a 2-fold increase in NOS3 levels favored by the incorporation of each C allele.NOS3 levels higher than 80% of the constitutive gene expression level (B2M) presented a 4-fold increase in PCa occurrence.The -786T>C polymorphism was the most important promoter alteration of the NOS3 gene that may affect the PCa progression, but not its occurrence, and the incorporation of the C allele is associated with increased levels of NOS3 transcripts.

View Article: PubMed Central - HTML - PubMed

Affiliation: Federal University of Uberlândia, Institute of Genetics and Biochemistry, Molecular Genetics Laboratory, Campus Umuarama, Block 2E, Room 24, 38400-902, Uberlândia, MG, Brazil. kmarangoni@yahoo.com.br

ABSTRACT

Background: There is no biological or epidemiological data on the association between NOS3 promoter polymorphisms and prostate cancer. The polymorphisms in the promoter region of NOS3 gene may be responsible for variations in the plasma NO, which may promote cancer progression by providing a selective growth advantage to tumor cells by angiogenic stimulus and by direct DNA damage.

Methods: This study aimed evaluating the NOS3 promoter polymorphisms by PCR-SSCP and sequencing, associating genotypes and haplotypes with NOS3 expression levels through semi-quantitative RT-PCR, and with PCA3 mRNA detection, a specific tumor biomarker, in the peripheral blood of pre-surgical samples from 177 patients; 83 PCa and 94 BPH.

Results: Three novel SNPs were identified -764A>G, -714G>T and -649G>A in the NOS3 gene promoter region, which together with the -786T>C generated four haplotypes (N, T, C, A). NOS3 gene expression levels were affected by the -786T>C polymorphism, and there was a 2-fold increase in NOS3 levels favored by the incorporation of each C allele. NOS3 levels higher than 80% of the constitutive gene expression level (B2M) presented a 4-fold increase in PCa occurrence.

Conclusion: The -786T>C polymorphism was the most important promoter alteration of the NOS3 gene that may affect the PCa progression, but not its occurrence, and the incorporation of the C allele is associated with increased levels of NOS3 transcripts. The NOS3 transcript levels presented a bimodal behavior in tumor development and may be used as a biomarker together with the PCA3 marker for molecular staging of the prostate cancer.

Show MeSH
Related in: MedlinePlus