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Associations have been found between the angiotensin-converting enzyme insertion deletion (I/D) polymorphism (ACE I/D) and endometrial and epithelial ovarian cancer (EC and EOC, respectively). In this study, the following frequencies for each of three ACE polymorphisms, DD, ID, and II, respectively, were observed: in the EC group, 55, 24, and 21% versus the control group 39, 40, and 21% (p = 0.033*); in the EOC group 49, 36, and 15% versus the control group 49, 33, and 18% (p = 0.82). According to these allelic distributions, DD carriers are 2.0 times more likely than individuals carrying the ID or II genotypes to develop EC; therefore, the DD genotype seems to be protective against EC. In contrast, no association was observed between ACE (I/D) polymorphism with EOC. The ACE (I/D) polymorphism might play a role in the pathogenesis of EC and it should be considered when identifying genetic markers for EC.

Cytokine receptors are associated with tumor cell growth by increasing proliferation, metastasis and regulating self-renewal of cancer stem cells (SCs). There is a strong association between cytokine IL-8 receptor (CXCR1) over-expression and cells displaying SC characteristics. Human chorionic gonadotropin (hCG) causes differentiation, inhibition of cell proliferation and increased apoptosis of the breast epithelium. hCG receptor (LHCGR) expression in breast tumors and in breast cancer cell lines is undetectable or low. In this study, our objective was to assess and compare the effects of hCG and a 15 amino acid hCG fragment of the hormone on mRNA expression of CXCR1 and LHCGR on normal breast epithelial cells (MCF-10F) by real time RT-PCR after treatment with hCG or a hCG fragment for 15 days. Cell proliferation was also measured. hCG and the hCG fragment decreased cell proliferation in both groups. The compounds upregulated LHCGR expression and downregulated CXCR1 expression. It is possible to postulate that an increase of LHCGR mRNA seems to respond to the decrease of CXCR1 expression. These genes probably act synergistically to reduce the amount of cancer SCs in the mammary gland. Thereby, the use of hCG or the hCG fragment as a therapeutic or preventive tool should be considered.

Background: Many types of cancer are associated with polymorphisms of the renin-angiotensin system. Our aim was to assess possible association between single-nucleotide polymorphisms (SNPs) of the angiotensin II receptor types 1 (A168G), and 2 (T1247G and A5235G) with breast cancer. Patients and Methods: 242 participating subjects were genotyped and allocated to case or control groups. Results: Genotype distribution (in %) was: for AGTR1 (A168G): AA, AG, GG = 61, 30, 09 for cases, and 69, 25, 06 for controls (p = 0.55); for AGTR2 (T1247G): TT, TG, GG = 84, 12, 04 for cases, and 81, 17, 02 for controls (p = 0.45); for AGTR2 (A5235G): AA, AG, GG = 32, 67, 01 for cases, and 53, 28, 19 for controls (p < 0.0001). Women carrying genotypes AA/AG in the intronic region of angiotensin II type 2 receptor had an 11-fold higher risk of breast cancer than GG carriers. Conclusions: Many types of cancer have been associated with polymorphisms of the renin-angiotensin system. For SNP A5235G, the GG genotype seems to be protective against breast cancer. The other 2 SNPs showed no association. However, SNPs T1247G and A5235G were associated with at least 1 clinical variable, with G being a predictor of better outcome. The use of SNPs A5235G and T1247G (the latter to a lesser degree) as genetic markers should be considered.

The Polycystic Ovary Syndrome (PCOS) is the most common androgenic disorder in women during reproductive life. PCOS may also be accompanied by metabolic syndrome and recent studies point to leptin as playing a role in disrupting infertility and in changing the energy balance in obese mice through its action on the hypothalamus. The aim is to assess the expression of the Polycomb & Trithorax Complexes genes in brain of mice transplanted with fat tissue from normal mice, in order to better understand the neuronal mechanisms underlying the reversion of PCOS. Three B6 V-Lepob/J mouse groups: Normal weight, obese and seven-day-treatment obese had their brain RNA extracted and submitted to an 84 Polycomb & Trithorax Complexes genes PCR Array plate and Metacore TM pathways localization. Genomic profiles obtained were compared to the ones of the normal-weight-mice group. Differentially expressed genes were 13% and 26% respectively to control and treatment. Major changes were in genes: Snai1/31; Smarca1/−17; Dnmt3b/4.7; Ezh1/ 15. Altered genes were associated to canonical pathways and provided 3 networks related to epi-* Corresponding author. E. H. da Silva Freitas et al. 178genetics. Underlying neuronal changes caused by leptin in obese mice brain, there is an important role being played by the histone code. Here there is evidence that leptin drives the chromatin packing to a more condensed pattern. Upregulation of methyltransferase genes, like Ezh1, favors this thought. In summary the Polycomb & Trithorax complexes might answer for the silencing of some downregulated genes in the obese mice brain when exposed to leptin.

Introduction: Renin Angiotensin System (RAS) components are not only related to breast cancer but also to other cancer types. Angiotensin-(1-7) [Ang-(1-7)] is an endogenous 7-amino acid peptide hormone of the renin-angiotensin system that has antiproliferative properties. The aim of this work is to analyze the action of Ang-(1-7) treatment in MCF-10F (normal) and in SKBR3 (tumoral) epithelial breast cells in their capacity to be droven to apoptosis, and also to assess the expression of the membrane receptors CD24 and CD44 in these cells. Methods: We used the SA Biosciences Human Apoptosis RT*2 Profiler PCR Array profiles to analyze the expression of 84 key genes involved in programmed cell death. Furthermore a flow cytometer (GUAVA) was used for quantification of the membrane proteins CD24 and CD44 in the MCF10F and in the SKBR3 breast cells after 24 hours of Angiotensin 1-7 stimulation. Results: Flow cytometric analysis using MCF10F cells showed that a 24h Ang1-7 stimulation period down regulates CD24 gene expression (about 43%) and it up regulates CD44 gene expression (about 16%). On the other hand there was no difference in the expression of both CD24 and CD44 proteins after Ang1-7 stimulation in SKBR3 tumoral cells. Furthermore, after the gene expression analysis by PCR Array, we found differential expression in MCF10F cells after Ang 1-7 treatment and we observed that the genes had their expression changed about 30% (2.1 to 514 fold increases) and 11% (2.6 to 3.8 fold decreases), respectively on a panel of 84 genes related to apoptosis. For SKBR3 cells, we found that the genes were altered in about 5.9% (2.6 to 5.4 fold increases) and 12% (2.1 to 3.3 fold decreases), respectively. Conclusion: Ang 1-7 appears to modulate the expression of membrane proteins CD24 and CD44 in MCF10F cells, also this hormone seems to induce apoptosis by inducing the expression of apoptosis related genes, such as, Abl1, Akt, Bax, Bcl2 (fold change, 3, 12, 6, and 6, respectively). Another important gene is the Hrk (harakiri, BCL2 interacting protein), that regulates apoptosis through interaction with death-repressor proteins Bcl-2 and Bcl-X(L), it displayed a fold change of 514. On the other hand in SKBR3 cells, comprised almost entirely by mammary stem cells, we found no regulation of CD24 and CD44 proteins by Ang1-7. Moreover, it was observed a very low differential expression in the genes related to the apoptosis pathway in these cells. However, the peptide Ang 1-7 seems to act preferentially in differentiated cells of the normal breast epithelium. Financial Support: FAPESP. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3031. doi:1538-7445.AM2012-3031

Introduction: Renin Angiotensin System (RAS) is correlated to breast cancer and to other types of cancer. Angiotensin I-7 [Ang-(I-7)] is an endogenous 7-amino acid peptide hormone of this system that has anti-proliferative properties. Interestingly, there are increasing evidences that RAS is implicated in the development of breast cancer. Objectives: We aimed to evaluate the expression of steroid hormone receptors (progesterone receptors (PR), estrogen receptors (ER), androgen receptors (AR), apoptosis and proliferation in tumoral (T47D) epithelial mammary cell line, after Ang-(I-7) treatment. Methods: Tumoral (T47D) cultured cell line was treated with angiotensin I-7 peptide and cells were harvested after 2, and 15 days, when expression analysis of steroid hormone receptors was performed by real time PCR. The same treatment was used for the apoptosis and cell cycle analyses, done using the 5HT-GUAVA flow cytometer. Results: Expression of steroid hormone receptors implicated to breast carcinogenesis such as genomic progesterone receptors (PR(A+B) and PRB), estrogen receptors (Era and ERb), androgen receptor (AR) were never investigated together in the sex hormone-sensitive human breast cancer cell T47D after angiotensin I-7 exposure. When T47D cells were treated with angiontensin I-7 peptide were all up-regulated of the control level (PRA+B (1.0-fold), PRB (2.3-fold), Era (7.8-fold), ERb (8.1-fold) and AR (1.9-fold) - p &lt; 0.01) in early treatment (2 days). However, important down-regulation of all receptors was observed after 15 days. T47D responded by down-regulating PRA+B (-3.5-fold), PRB (-3.1-fold), Era (-4.3-fold), ERb (-4.6-fold) and AR (-6.05-fold), all compared to basal conditions (p &lt; 0.001). The functional relevance of steroid hormone receptors inhibition in sensitive human breast cancer cell line was confirmed by flow cytometry, assaying cell viability, apoptosis and proliferation by flow cytometry. Ang-(I-7) 2 day treatment significantly increased cell cycle in all phases (G0/G1, S and G2/M) when compared to control and decreased apoptosis (p &lt; 0.05), but after 15 days there was a significant reduction in all cell cycle phases (p &lt; 0.05) and increased apoptosis indexes were observed (p &lt; 0.001). Conclusion: These results suggest anti-proliferative actions of Ang-(I-7), which has already been demonstrated previously for this peptide by our research group (Alecrim et al, 2011). Moreover, we could also observe a pro-apoptotic effect sustained for 15 days in these cells, and it seems to be correlated to the down-regulated expression of all steroid hormone receptors studied. In summary, Ang-(1-7) drives T47D cells to apoptosis and the use of the peptide as a novel therapeutic compound for breast cancer should be considered. Support: FAPESP Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 266. doi:1538-7445.AM2012-266

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