Introduction: Claudins are tight junction molecules that have been associated with breast cancer prognosis. The claudin-low intrinsic subtype of invasive carcinoma was recently described and associated with high grade carcinoma, low junction molecules expression and worse response to chemotherapy. However, it is not known whether the expression of claudins may provide clues as to carcinoma in situ prognosis. The aim of study was evaluate the association between claudin–4 expression and disease-free survival and histologic type of local recurrence in carcinoma in situ after longer follow up. Methods: A tissue microarray (TMA) block was constructed from 137 pure carcinoma in situ paraffin blocks sampled from patients treated from 1999 to 2009. The TMA was submitted to immunohistochemical staining for claudin-4. A claudin-4 score calculated based on percentage and intensity of expression, categorized samples as: claudin-4-low or claudin–4-high. Clinical data, treatment data, local recurrence data and survival of each patient were reanalyzed from medical records. Kaplan-Meier curves, log-rank and Wilcoxon tests were used to analyze disease-free survival; qui-square and Fisher test were employed to compare others variables; a significance level of 5 % was used. Results: Claudin-4 expression was evaluated in 86 samples, 88.4% were claudin-4-high and 11.6% claudin-4-low. Mean follow up was 8.2 years ( and local recurrence rate was 10.5 %. There was significant difference in the disease-free survival between claudin-4-high and claudin-4-low (4.9 x 1.9 respectively, p= 0.02); however there was no difference between both in histologic type of recurrence, invasive or in situ (p=0.44). Conclusion: In our samples, claudin-4-high expression in carcinoma in situ was more frequent than low expression. Our data showed that claudin-4-low expression had a worse prognosis in carcinomas in situ (inferior disease-free survival) but it was similar to claudin-4-high in histologic type of local recurrence. Citation Format: Duarte GM, Almeida NR, Tocchet F, Espinola J, Pinto T, Barreto CT, Pinto GA, Soares FA, Marshall P, Paiva GR. Claudin-4 expression is associated with disease free survival in breast carcinoma in situ: Mean follow up of 8.2 years [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P3-17-05.

Introduction: Claudins are tight junction molecules and have been associated to breast cancer prognosis. Claudin-low intrinsic subtype of invasive carcinoma was described recently and has been related to high grade carcinoma, low junction molecules expression and worse chemotherapy response. However, it is unknown whether Claudins expression could be associated to carcinoma in situ prognostic. The aim of this study was evaluated the Claudin – 4 expression in carcinoma in situ and its association with local recurrence, clinical and immunohistochemistry characteristics. Methods: A tissue microarray (TMA) block was constructed, using region of interesting, with 137 pure carcinoma in situ paraffin blocks of patients treated in the Women 's Hospital Prof. Dr. José Aristodemo Pinotti – UNICAMP from 1999 to 2009. The TMA was submitted to immunohistochemistry analyze to: Claudin-4, beta-catenin, e-caderin, estrogen receptor (ER), progesterone receptor (PR), HER-2 and Ki-67. It was calculated Claudin-4 score based in percentage and intensity of expression and categorized in: Claudin-4 low and Claudin – 4 high. The clinical data, treatment data (surgery, radiotherapy and tamoxifen use), local recurrence data (date and type) and death of each patient were reviewed in the medical records. The statistical analyze used Kaplan-Meier curve and log-rank test to disease free survival; qui-square and Fisher test to compare others variables; significance level of 5 % was used. Results: It was possible to evaluate Claudin-4 expression in 86 cases, 88.4% were Claudin-4 high and 11.6% Claudin-4 low. The follow up mean was 69 months and local recurrence rate was 10.5 %. There was no significant difference in local recurrence rate between Claudin-4 high and Claudin-4 low (10.0% x 10.5% , p=1.0).The disease free survival was similar between Claudin-4 low and Claudin-4 high (p=0.559). The Claudin- 4 high was significantly more frequent in beta-catenin positive patients (p=0.048). There was no association significantly between Claudin-4 expression and: age (p=0.66), histology type (p=0.75), surgery (p=0.102), radiotherapy (p=0.29), tamoxifen use (p=0.432), ER (p=0.33), PR (p=1.0), HER-2 (p=0.23) and e-caderin (p=0.21). Conclusion: Despite the Claudins are related to invasive carcinoma prognosis, our outcome did not show difference in local recurrence and disease free survival between Claudin-4 low and high in carcinoma in situ. The beta-catenin and claudin-4 expressions were significantly associated. Citation Format: Duarte GM, Toucchet F, Espinola JP, Barreto CR, Paiva Silva GR, Almeida NR, Soares F, Pinto G, Marshall P. Claudin -4 expression in carcinoma in situ and its association with local recurrence, clinical and immunohistochemistry characteristics. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P5-17-10.

Objective: We investigated immunohistochemical and histological composition of dense and non-dense breast tissue in 18 women undergoing mastectomy as the initial treatment for breast cancer. Materials and Methods: In each mammogram, we localized the dense and the non-dense areas. We used a localization technique based on a linear approximation method with interpolation of mammogram images and breast pictures. The selected areas were retrieved during mastectomy and analyzed. Results: Estrogen and progesterone receptors, Ki-67 and CD-34, were equally expressed in both tissues, as well as the percentage composition of fat. The percentage compositions of brownish spots among dense and non-dense tissues were significantly different (p = 0.0226). The number of terminal ductal lobular units was higher for dense than for non-dense breast tissues (p = 0.0019). In the non-dense breast tissue, there were no proliferative lesions with atypia, while we found flat epithelial atypia in 3 of the dense areas evaluated. Proliferative lesions without atypia and non-proliferative lesions were found in both tissues, but they were more frequent in dense than in non-dense breast tissues (23.5% vs 11.8%, p = 0.0455, and 17.6% vs 2.9%, p = 0.0253, respectively). Fibrosis was more frequently extensive or moderate in dense tissue, while it was predominantly mild in non-dense tissue (p = 0.03). Conclusion: There was no difference in the expression of the estrogen and progesterone receptors, Ki-67 and CD-34, in the dense and non-dense tissue areas in breast-cancer women. In addition, both stroma fibrosis and epithelial proliferation were responsible for higher mammographic density. Citation Format: dos Santos CC, Marshall P, Torresan R, Tinóis E, Duarte G, Teixeira S. Immunohistochemical and histological features of mammographic dense and non-dense tissue in breast cancer patients. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P4-01-04.