Ana Belén Peinado Lozano

DETECTION OF BREAST CANCER CELLS IN THE PERIPHERAL BLOOD IS POSITIVELY CORRELATED WITH ESTROGEN-RECEPTOR STATUS AND PREDICTS FOR POOR PROGNOSIS

We investigated whether detection of cytokeratin‐positive (CK+) cells in the peripheral blood (PB) of breast cancer patients before chemotherapy could be a prognostic factor. Blood from a total of 92 breast cancer patients was evaluated for the presence of CK+ cells. Blood samples were collected before chemotherapy. Patients entered in the study included: neoadjuvant (n = 25), adjuvant (n = 42) and metastatic (n = 25). Blood samples (10 ml) were centrifuged using a double density‐gradient to recovering the mononuclear cell (MNC) and granulocyte cell (GC) fractions. Subsequently, positive immunomagnetic cell separation was carried out to isolating CK+ cells. The enriched cell fraction was cytocentrifuged and then immunocytochemically labeled using an anti‐cytokeratin antibody. Our results indicated that breast tumor cells sediment with both MNC and GC fractions. We therefore recommend examination of both fractions in all enrichment protocols. CK+ cells in PB were identified in 57 of 92 (62%) patients when MNC and GC fractions were assessed (range = 1–61 cells, median = 8). No CK+ cells were detected in blood samples of 16 healthy donors. There were significant differences in the presence of CK+ cells according to estrogen receptor expression (p = 0.049), and lymph node status (p = 0.033), but not to the age, menopausal status, type of patient (neoadjuvant, adjuvant or metastatic), TNM stage, histological type, progesterone receptor expression, c‐erbB2 expression, p53 expression or Ki67 expression. Regarding the relationship between tumor size (T) and the presence of CK+ cells, a borderline significant trend was observed (p = 0.07). The median follow‐up of the patients was 21 months and statistical analysis (Kaplan‐Meier analysis) showed that using the method we present, the detection of CK+ cells in PB before starting the chemotherapy in breast cancer patients was significantly correlated with both progression‐free survival (p = 0.058) and overall survival (p = 0.003). In conclusion, the present study suggests that detection of CK+ cells in PB before chemotherapy might identify breast cancer patients with poor prognosis.

Evaluation of a Gemcitabine-Doxorubicin-Paclitaxel Combination Schedule through Flow Cytometry Assessment of Apoptosis Extent Induced in Human Breast Cancer Cell Lines

Combination chemotherapy with gemcitabine (Gem), doxorubicin (Dox), and paclitaxel (Pac) (GAT) has been considered attractive as first‐line treatment in metastatic breast cancer. We compared the potential of various schedules of GAT to induce apoptosis on MDA‐MB–231, MCF7, and T47D human breast cancer cell lines. The extent of apoptotic induction was analyzed by flow cytometry with 7–aminoactinomycin D (7AAD) staining. Differences between various schedules in terms of apoptotic induction were statistically significant (P<0.05). The most effective apoptotic induction regimen was achieved by the sequence: Dox for 16 h followed by Pac+Gem. Schedules employing a 16–h interval between drug administrations induced higher levels of apoptosis in human breast cancer cell lines compared with schedules using a 4–h interval. The therapeutic efficacy of the experimental results shown in this paper has been clinically corroborated in a phase II trial in metastatic breast cancer patients.

Lipid profile response to weight loss program in overweight and obese patient is related with gender and age.

INTRODUCTION The risk of developing cardiovascular disease (CVD) increases with age, however CVD is markedly higher in men than in no-menopausal women. There are few interventions where compare the different effects to lose weight on lipid profile between men and women. OBJECTIVE The aim of this study was to compare the response on the lipid profile by gender after a weight loss program, and determine whether there are differences by age group. METHODS One hundred eighty (96 women and 84 men) overweight and obese participants (BMI 25-34.9 kg/m2) aged 18-50 years were randomised into treatment groups. The intervention period was 22 weeks (in all cases 3 times/wk of training for 22 weeks and 2 weeks for pre and post evaluation). All subjects followed a hypocaloric diet (25-30% less energy intake). Energy intake, body composition) and blood lipid profile were recorded at baseline and after of treatment. RESULTS The response of HDL varied between men and women (p = 0.001). While in women it decreased (HDL: -2.94%, p = 0.02), HDL was elevated in men (HDL: 5% p = 0.02). After intervention men achieved decrease significantly LDL values a 6.65% more than women (p = 0.01). For TG concentrations there were significant differences between men and women in baseline however, only men had a significant chance in post-training measured (p = 0.001). TC showed significant differences between men and women in baseline (p = 0.013). After intervention, men and women showed a significant decreased to TC (p = 0.01). CONCLUSION Men achieve a positive greater change on lipid profile than women. In addition, the favorable lipid profile response decreases with increasing age.