Ana Maria Massad Costa

Association between the angiotensin-converting enzyme (insertion/deletion) and angiotensin II type 1 receptor (A1166C) polymorphisms and breast cancer among Brazilian women

Introduction.We evaluated the association between components of the renin-angiotensin system and the development of breast cancer in a case-control study by means of angiotensin-converting enzyme (ACE) insertion/deletion (I/D) and angiotensin II type 1 (AT 1)-receptor A1166C polymorphisms. Methods. Genotyping was performed by PCR-RFLP (restriction fragment length polymorphism) or PCR (polymerase chain reaction) using genomic DNA extracted from buccal cells of subjects with (101 cases) or without (307 controls) breast cancer. Results.The frequencies of genotypes for ACE were: DD, ID and II (in %: cases: 60; 20; 20; controls: 46; 37; 17; p=0.019, χ2); and for AT1receptor were:AA,AC and CC (in %: cases: 65; 30; 5; controls: 51; 44; 5; p=0.114, χ 2).The results suggested that the A1166C polymorphism was not associated with breast cancer risk. On the other hand, for the ACE (I/D), there seemed to be different risks for cancer between cases and controls. Conclusions.The ID genotype was less frequently associated with the disease than were the DD or II; that is, women with the ID genotype were 3.1 times less likely to develop breast cancer than those with the other genotypes.The ID genotype might be protective against breast cancer and the ACE (I/D) polymorphism a possible target for developing genetic markers for breast cancer.

Genetic polymorphisms of cytochrome P450c17α (CYP17) and progesterone receptor genes (PROGINS) in the assessment of endometriosis risk

We designed the present study in order to evaluate the eventual role of polymorphisms in the genes encoding cytochrome P450c17α (CYP17) and the progesterone receptor (PROGINS) as risk factors for endometriosis development. Eligible cases consisted of 121 women with surgically confirmed endometriosis who underwent treatment in a hospital in São Paulo, Brazil during the period from September 2003 to September 2005. The 281 controls were participants with normal gynecological as well as pelvic ultrasound evaluation, who did not have any gynecological conditions during their reproductive lives such as pelvic pain and/or dyspareunia nor infertility history. Genomic DNA was obtained from buccal cells and processed for DNA extraction using the GFX DNA extraction kit (GE Healthcare). The CYP17 (−34T→C) polymerase chain reaction–restriction fragment length polymorphism assay has been described previously, as has the progesterone receptor polymorphism (PROGINS) detection assay. PROGINS heterozygosis genotype frequencies were shown to be statistically higher in endometriosis cases compared with controls. On the other hand, differences in the CYP17 polymorphism (−34T → C) frequencies were not even close to significance (p = 0.278) according to our findings.

Association between estrogen receptor gene polymorphisms and breast density in postmenopausal women

Objectives The aim of this study was to evaluate the association between clinical characteristics and polymorphisms HaeIII, MspI and XbaI of the estrogen receptor gene α with postmenopausal mammographic density. Methods A prospective study was performed with 120 women who were not users of hormones and had no identified breast lesions. All of them underwent bilateral mammography; the radiological density was determined by three independent observers, with two subjective evaluations based on the ACR-BIRADS® classification of mammographic patterns, 2003, and one computerized evaluation using the gray-scale histogram tool of the Adobe Photoshop® 7.0 software. Peripheral blood samples were obtained for DNA extraction, performed according to the GFX® Kit protocol (Amersham-Pharmacia). Polymerase chain reaction restriction fragment length polymorphism was carried out for an analysis of the polymorphisms present in intron 1 (HaeIII and XbaI) and in exon 1 (MspI) of the estrogen receptor gene. Results There was a high degree of concordance among the observers in the determination of mammary density (Kappa, Pearson and Spearman, p < 0.001). The associations of clinical characteristics with mammary density were: age (p = 0.04), body mass index (p < 0.0001) and age at menarche (p = 0.02). The relationship between the allele distribution of the polymorphisms and density was: XbaI (p = 0.02), HaeIII (p = 0.65) and MspI (p = 0.65). Conclusions Our data suggested that the polymorphism XbaI may be strongly related to mammographic density.

Polymorphism in CYP17, GSTM1 and the progesterone receptor genes and its relationship with mammographic density

Radiologic breast density is one of the predictive factors for breast cancer and the extent of the density is directly related to postmenopause. However, some patients have dense breasts even during postmenopause. This condition may be explained by the genes that codify for the proteins involved in the biosynthesis, as well as the activity and metabolism of steroid hormones. They are polymorphic, which could explain the variations of individual hormones and, consequently, breast density. The constant need to find markers that may assist in the primary prevention of breast cancer as well as in selecting high risk patients motived this study. We determined the influence of genetic polymorphism of CYP17 (cytochrome P450c17, the gene involved in steroid hormone biosynthesis), GSTM1 (glutathione S-transferase M1, an enzyme involved in estrogen metabolism) and PROGINS (progesterone receptor), for association with high breast density. One hundred and twenty-three postmenopausal patients who were not on hormone therapy and had no clinical or mammographic breast alterations were included in the present study. The results of this study reveal that there was no association between dense breasts and CYP17 or GSTM1. There was a trend, which was not statistically significant (P = 0.084), towards the association between PROGINS polymorphism and dense breasts. However, multivariate logistic regression showed that wild-type PROGINS and mutated CYP17, taken together, resulted in a 4.87 times higher chance of having dense breasts (P = 0.030). In conclusion, in the present study, we were able to identify an association among polymorphisms, involved in estradiol biosyntheses as well as progesterone response, and radiological mammary density.

Positive association of the hepatic lipase gene polymorphism c.514C > T with estrogen replacement therapy response

BackgroundHepatic lipase (HL), an enzyme present in the hepatic sinusoids, is responsible for the lipolysis of lipoproteins. Human HL contains four polymorphic sites: G-250A, T-710C, A-763G, and C-514T single-nucleotide polymorphism (SNPs). The last polymorphism is the focus of the current study. The genotypes associated with the C-514T polymorphism are CC (normal homozygous - W), CT (heterozygous - H), and TT (minor-allele homozygous - M). HL activity is significantly impaired in individuals of the TT and CT genotypes. A total of 58 post-menopausal women were studied. The subjects were hysterectomized women receiving hormone replacement therapy consisting of 0.625 mg of conjugated equine estrogen once a day. The inclusion criteria were menopause of up to three years and normal blood tests, radiographs, cervical-vaginal cytology, and densitometry. DNA was extracted from the buccal and blood cells of all 58 patients using a commercially available kit (GFX® - Amersham-Pharmacia, USA).ResultsStatistically significant reductions in triglycerides (t = 2.16; n = 58; p = 0.03) but not in total cholesterol (t = 0.14; n = 58; p = 0.89) were found after treatment. This group of good responders were carriers of the T allele; the CT and TT genotypes were present significantly more frequently than in the group of non-responders (p = 0.02 or p = 0.07, respectively). However, no significant difference in HDL-C (t = 0.94; n = 58; p = 0.35) or LDL-C (t = -0.83; n = 58; p = 0.41) was found in these patients.ConclusionsThe variation in lipid profile associated with the C-514T polymorphism is significant, and the T allele is associated with the best response to ERT.

Abstract 5592: Lack of association between AGTR1 (A168G and T825A) and ACE T5529C polymorphisms with breast cancer among Brazilian women

Background: Angiotensin II (AngII) exerts promitotic, pro-proliferative and angiogenic effects and the administration of angiotensin I-converting enzyme (ACE) inhibitors reduces tumor growth in animal models of cancer. AngII type 1 receptor (AGTR1) is found in a wide variety of normal tissues, increased expression is often found in the corresponding neoplastic tissues, suggesting that its overexpression is involved in carcinogenesis. In a previous study we observed that the ID genotype might be protective against breast cancer and also that the ACE (I/D) polymorphism is a possible target for developing genetic markers for breast cancer. Other authors have also observed the association of the ACE with breast cancer risk. To further test the hypothesis that AngII participates in breast carcinogenesis through AGTR1 and ACE, we examined genetic polymorphisms in the 5’-region of the AGTR1 gene (A-168G and T-825A) and in the ACE (T5529C) in relation to risk of breast cancer among Brazilian women. Methods: Genotyping was performed through Real Time PCR (A168G and T825A) or PCR-RFLP (T5529C) using genomic DNA extracted from buccal cells of subjects with or without breast cancer.Results: Patients with (case) or without (control) breast cancer aging from 30 to 90 years old were genotyped for A168G (n=516), and the following frequencies obtained for AA, AG and GG in % were, among cases: 63, 30, 07 and among controls: 64, 32, 04 (p = 0.89). For T825A (n=562), we determined the frequency of TT, TA and AA (in %, cases: 62, 33, 05; controls: 59, 33, 08; p = 0.68). At last, for ACE (T5529C) (n=272) these are the obtained frequencies for TT, TC and CC genotypes in %, among cases: 65, 23, 12 and among controls: 55, 29, 15 (p = 0.39). Conclusion: Our results suggest a lack of significant association between AGTR1 polymorphisms (A168G and T825A) with breast cancer risk in the Brazilian population, in contrast to a previous result among Chinese women, where an association of the AGTR1 polymorphisms with breast cancer has been reported. Regarding the ACE T5529C polymorphism, it also did not show any association with breast cancer risk in the present study, differently from the observed for another polymorphism of the same gene, the ACE (I/D). Therefore, the AGTR1 (A168G and T825A) and ACE (T5529C) variant genotypes do not appear to be related to risk of breast cancer among Brazilian women. Support: FAPESP. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5592. doi:10.1158/1538-7445.AM2011-5592

Association of estrogen receptor gene alpha polymorphisms with postmenopausal mammary density.

Abstract #5003 Background: With the human genome studies, knowledge about polymorphisms started raising interest in a variety of fields and, in medicine, the evidence of direct action of polymorphisms on the arising and progression of diseases, disclosing the possibility of using them as disease predisposition markers. Substitutions, insertions or deletions which are transmitted through generations and reach frequencies equal or superior to 1% in the population are named polymorphisms. Knowing that the mammographic pattern is a multifactorial character, the objectives of this study were to evaluate a possible association of clinical characteristics and polymorphisms HaeIII, MspI and XbaI of the estrogen receptor gene alpha with postmenopausal mammary density. Materials and Methods: A prospective evaluation was made of 120 women who were not hormone therapy users and had no clinically or mammographically identified breast lesions. All of them underwent bilateral mammography, and the radiological density was determined by three independent observers, with two subjective evaluations based on the ACR-BIRADS ® classification of mammographic patterns, 2003, and one computerized evaluation – the grey-scale histogram tool of the Adobe Photoshop ® 7.0 software. Peripheral blood samples were obtained for DNA extraction, performed according to the GFX ® Kit protocol from Amersham-Pharmacia. After DNA extraction, PCR-RFLP (Polymerase Chain Reaction - Restriction Fragment Length Polymorphism) was carried out for an analysis of the polymorphisms present in intron 1 (HaeIII and XbaI) and in exon 1 (MspI) of the estrogen receptor gene. Results: There was a high degree of concordance among the observers in the determination of mammary density (Kappa, Pearson and Spearman - p Conclusion: Polymorphism XbaI and the clinical factors age, menarche and body mass index showed to be associated with postmenopausal mammary density. Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 5003.