Altered levels of plasma chemokines in breast cancer and their association with clinical and pathological characteristics.

Chemokines are a family of small, structurally related cytokines with chemoattractant and activation properties. In breast cancer, both epithelial cancer cells and cells within the microenvironment secrete chemokines with either tumor-promoting or anti-malignant potential. The equilibrium between these two chemokine activities plays a key role in the biology of the developing tumor, including its ability to metastasize. Here we evaluated the expression of chemokines in breast tumors and the plasma of breast cancer patients before treatment in order to identify a blood-based signature that could distinguish between malignant and non-malignant processes. We screened the mRNA expression of chemokine genes using cDNA microarray on homogenous, laser-capture microdissected breast cancer specimens. Further, using a protein array approach, we determined the levels of selected chemokines in the plasma of patients with breast cancer, benign breast tumors and healthy women. Finally, we analyzed the association between the levels of chemokines in breast and blood samples with the pathological characteristics of the disease. At mRNA level, 27 chemokines and 11 chemokine receptors were differentially expressed in cancers when compared with normal breast tissue. When compared to benign tumors, the only chemokine significantly upregulated in cancers was CXCL10. At protein level, with the exception of CXCL13, nine out of the ten selected chemokines (CCL2, CCL7, CCL18, CCL22, CXCL8, CXCL9, CXCL10, CXCL11 and osteoprotegerin) were significantly overexpressed in the plasma of breast cancers patients compared to healthy controls. After grouping, CXCL8, CXCL9 and CCL22 proved to be significant predictors for breast cancers as compared to healthy controls in a model of logistic regression. We found upregulation of CXCL8, CXCL11 and CXCL9 in triple negative carcinomas, CXCL9 in low proliferative carcinomas, and CXCL10, CCL7 and osteoprotegerin in poorly differentiated carcinomas. Furthermore, CXCL9 was overexpressed in lymph node negative tumors, whereas CXCL8 and CCL18 were higher in advanced stage carcinomas. We identified a panel of chemokines dysregulated in breast cancer that could be further investigated as prospective novel diagnostic markers or for therapeutic and prognostic applications.

Treatment of complete genital prolapse by sacrospinous fixation, anterior mesh repair and conservation of the uterus.

UNLABELLED: The surgical treatment of complete genital prolapse must aim the restoration of the vaginal support structures. All 3 levels of the vaginal support system must be recreated. Ablation of the uterus is not useful for pelvic floor support. Sparing of the uterus offers the advantage of a reduced surgical trauma, and better pelvic floor restoration. We present the case of a 60-year old woman with complete genital prolapse where the uterus was spared, and sacrospinous fixation, anterior mesh repair,perineal body repair and suburethral sling insertion were performed. The results were very good, by means of pelvic floor statics and physiology of micturition. CONCLUSION: genital prolapse must be cured by reconstruction of the vaginal support system, not by hysterectomy.

Expression of CCL18 and interleukin-6 in the plasma of breast cancer patients as compared with benign tumor patients and healthy controls.

A growing body of laboratory research has shown that pro-inflammatory cytokines can facilitate tumor growth and metastasis. Our goal was to quantify the expression of CCL18 and IL-6 in patients with breast cancer compared with benign breast tumors patients and healthy women, in order to evaluate if these cytokines could serve for breast cancer diagnosis and evaluation. We also correlated the cytokines level of expression with some clinical and pathological characteristics known as prognostic markers for breast cancer. Plasma samples were obtained before treatment from 58 breast cancers, 41 benign breast tumors and 30 healthy women. The quantitative dosage was performed using ELISA. Wilcoxon test was used to compare groups. IL-6 and CCL18 were dramatically upregulated in breast cancers in comparison with healthy controls, but in comparison with benign tumors only CCL18÷PARC was overexpressed at borderline significance in cancers (p=0.05). The plasma from benign breast tumor patients exhibited also significant higher levels of the two cytokines than normal controls. The cytokines profile was not linked to patient age, tumor size, histopathological type, lymph node status or histological grade. IL-6 was significantly upregulated in ER-positive and metastasized cancers. CCL18÷PARC presented a significantly higher expression in advanced stage and highly proliferative carcinomas. In summary, IL-6 and CCL18 could clearly distinguish between women with breast cancers and healthy controls. High expression of IL-6 seems to confer a poor prognosis for ER-positive cancers. CCL18 was associated with worse prognosis parameters like high Ki67.

Molecular profiling of ADAM12 gene in breast cancers.

ADAMs (a disintegrin and metalloproteinase) family have been associated with the process of proteolytic "shedding" of membrane-associated proteins ectodomain and hence the rapid modulation of key cell signaling pathways in tissues microenvironment. A variety of cytokines, chemokines and growth factors which are initially produced as transmembrane proforms are activated by these sheddase activities. ADAM12 is highly expressed in rapidly growing tissues such as placenta and malignant tumors and it was found as one of the Candidate Cancer Genes in a comprehensive mutational analysis of human breast cancers. Our aim was to determine the gene expression profile of ADAM12 in breast cancers in comparison with normal breast and to correlate their level of expression with the clinical and pathological characteristics of breast cancers. Gene expression of ADAM12 spliced variants (12L and 12S) was evaluated using quantitative reverse-transcription PCR in samples obtained by laser capture microdissection from 38 patients with breast cancers and compared with adjacent healthy breast tissues. Both ADAM12L and 12S expression were significantly up-regulated in breast cancers, while in the normal breast, we found a very low expression. ADAM12L expression was significantly correlated with the histopathological types and, although not statistically significant, ADAM12 both variants were up-regulated in high-grade, highly-proliferative and HER2÷neu positive tumors. From these preliminary results, we found that ADAM12 could be an interesting marker and eventually a therapeutic target for breast cancer.

Interaction between estrogens and androgen receptor genes microsatellites, prostate-specific antigen and androgen receptor expressions in breast cancer.

The role of estrogen and androgen receptors signaling in breast cancer is widely accepted, but the interrelations between them are not well understood. It was suggested that PSA could be a marker of endogenous balance between androgens and estrogens. In this context, we intended to investigate the potential of relationship between polymorphic tandem repeats (CAG, TA and CA) in AR (androgen receptor), ERalpha (estrogen receptor alpha) and ERbeta (estrogen receptor beta) genes and the immunoexpression of PSA and AR proteins. We assessed also the possible influences of CAG, TA, and CA variables and other available prognostic factors (ER, PR, AR, HER2/neu, PSA expression, and nodal status) on disease-free survival. We assessed the polymorphic tandem repeats lengths by genotyping, followed by high-resolution denaturing polyacrylamide gel electrophoresis in 163 breast cancers. Immunohistochemistry was performed to assess the expressions of AR, PSA, ER, PR and HER2/neu proteins. Our results showed that PSA was correlated with the length of CA repeats in the 3'-untranslated region of ERbeta, shorter CA repeats being correlated with PSA expression (p=0.03). AR immunoexpression was correlated with CAG repeats on AR gene, higher number of repeats being linked to a higher AR immunoexpression (p=0.04). Performing logistic regression to investigate relationships with prognosis, we observed that PSA immunoexpression (p=0.004), the nodal status (p-<0.001) and marginally, longer TA repeats (p=0.05) were correlated with increased disease-free survival. AR expression presented a low statistical value (p=0.054) in predicting evolution and was not entered into the multivariate regression analysis. Altogether, our findings supports the hypothesis that estrogens, through both alpha and beta-receptors variants are mediating the AR signaling pathway.

Role of multi-component functional foods in the complex treatment of patients with advanced breast cancer.

UNLABELLED: Cancer increases free radicals and that certain diets can influence oxidative stress. There is little evidence about the functional effect of multi-component functional foods on oxidative stress in breast cancer. MATERIAL AND METHODS: 34 female patients with histologically confirmed breast cancer, stages IIIB and IV were enrolled in a randomised controlled trial. All patients filled out the Quality of Life questionnaire FACT-B version 4, translated in Romanian and validated, obtained from Evanston North-Western Health Care, Illinois, USA. Seventeen patients, mean age 57.84 +/- 11.2, were put on a special diet consisting of 15 ml/day balsamic vinegar from apples and honey, with seabuckthorn berry rich in carotenoids, rosemary, sage and basil extracts, to be used in salads and vegetable soups, 150 g/day whole wheat bread with 2.5% of the nutraceutic mixture VITAPAN, 15 ml/day grape seed extract rich in polyphenols (gr. 1). Seventeen patients served as control (gr. 2). Parameters of the metabolic syndrome were measured: serum lipids, IR-HOMA insulin resistance index. Oxidative stress was measured by the FORT (free oxygen radical) test. Total hydro- and liposoluble antioxidants (ACW, ACL) in serum were measured by chemo-luminometry. All investigations were done at entry and after 3 months. RESULTS: Student's t test was applied to compare the means within group and between the 2 groups. The Physical well being subscale score of the QOL FACT-B version 4 questionnaire showed a significant difference between the 2 groups (p = 0.001). Also, functional well being subscale (p = 0.004), FACT-G (p = 0.003) and FACT-B (p = 0.002) showed significant differences on comparison. Breast Cancer Score (Additional Concerns) had a borderline significant difference (p = 0.057). Social/Family Well being subscale and Emotional well being subscale scores showed no significant difference. At inclusion, radical activity > 310 FORT units, relevant for increased oxidative stress were present in 95.1% cases. After 3 months radical activity > 310 FORT units were present in 52.8% cases in gr. 1. CONCLUSION: A proper design of foods enriched with bioactive compounds that can lead to significant effects on health represents a promising adjuvant treatment in patients with advanced breast cancer, due to its contribution in lowering the high oxidative stress present in these patients.