- Email: [email protected]
1898 Characteristics of the local immunity of breast cancer patients with different reproductive function statuses
S298 Fibroblasts (cCAFs) either as markers and predictors of metastasis or as a prognostic indicator in patients with MBC has not been studied. The purpose of this study is to demonstrate the presence of cCAFs simultaneously with CTCs in patients with MBC. Material and Methods: A total of 7.5 ml of peripheral blood from each of twenty patients with MBC (Metastatic group) and six patients with cured Breast Cancer (DCIS or Stage I post definite treatment with greater than five years of disease free survival, Localized group) was collected. The micro filter capture process was used to analyze CTCs and cCAFs. Utilizing a triple immufluorescence staining for pan-CK (cytokeratin), FAP (Fibroblast Activated Protein) and CD45, cCAFs were identified as CK−, FAP+, CD45− cells whereas CTCs were identified as CK+, CD45− cells. cCAF identification was also confirmed in parallel samples by a FAP/a-Smooth Muscle Actin staining. Results: cCAFs were detected in 16/20 (80%) of patients from the Metastatic group and in 1/6 (16.7%) of the patients from the Localized group. Likewise, CTCs were detected in 20/20 (100%) patients from the Metastatic group and in 5/6 (83.3%) from the Localized group. The counts of CTCs and cCAFs in the Metastatic group ranged between 1−98(median 12) and 2–117(median 4), respectively. The counts of CTCs and cCAFs in the Localized group ranged between 1−10(median 6) and 0−1 (median 0) respectively. Conclusions: A sharp contrast of CTC and cCAF levels was observed between the Metastatic and the Localized groups. These findings associate CTCs and cCAFs with advanced stage disease, and support cCAFs as a promising biomarker for metastatic breast cancer. We have successfully, for the first time, demonstrated that cCAFs can be enumerated in patients with metastatic breast cancer alongside with CTCs. The interaction between CTCs and cCAFs and their role in metastasis requires additional investigation. Validation of these findings in a larger cohort will be presented during the meeting. Conflict of interest: Ownership: Dr Ram Datar is among the inventors of intellectual property for CTC microfilters, which are used in the study, and may gain royalties. from future commercialization of the microfilter technology.
Poster Session (Monday, 28 September) Breast Cancer − Early Disease 1896 POSTER Correlation between body mass index (BMI) and histopathologic characteristics of breast cancer and recurrence rate in pre and postmenopausal women W. El-Sadda1 , I. Abdel-Halim2 , M. Magdy2 , M. El-Ibrashy2 , H. Daghriri1 . 1 Al-Ghad International Colleges for Health Sciences, Radiology, Najran, Saudi Arabia; 2 Mansoura University Hospital, Clinical Oncology & Nuclear Medicine, Mansoura, Egypt Background: High BMI is related to increased breast cancer incidence to more aggressive and poorer prognosis, The study aims to analyze the association between increased body weight by using the body mass index (BMI) at time of diagnosis and histopathologic features of breast cancer (tumor size, nuclear grade, hormone receptors (ER & PR), HER-2/neu expression, Ki-67 index, lymphatic/vascular invasion, axillary nodes involvement and menopausal status, to evaluate the impact of BMI on DFS at multivariate analysis. Patients and Methods: In this retrospective study, 500 patients with early breast cancer (200 premenopausal and 300 postmenopausal) were classified into subgroups according to BMI; normal was defined as BMI <25 kg/m2 , overweight as BMI >25 kg/m2 and obese as BMI >30 kg/m2 . The association between BMI and histopathologic features, ki-67 labeling index was evaluated. Results: High BMI significantly associated with larger size tumor both in premenopausal (p = 0.01) and postmenopausal women (p = 0.001) Obese premenopausal women were associated with more aggressive histopathologic characteristics (e.g., more positive axillary nodes p = 0.016 and presence of vascular invasion p = 0.005) compared to normal weight group. Postmenopausal patients with BMI >25 developed more frequently ER/PR positive cancer (85% vs. 70%, p = 0.01) while no association was found in premenopausal women. BMI did not show significant association to tumor nuclear grade, HER-/neu overexpression, Ki-67 index and lymphatic invasion in both pre and postmenopausal women. In multivariate analysis, obesity was not an independent prognostic factor in pre and postmenopause, however, obese women showed a shorter DFS and in the postmenopausal group with BMI >25 as a single variable had a statistically significant worse DFS compared to normal weight women.
Abstracts Conclusion: Obesity plays a different role in breast cancer according to menopausal status; postmenopausal women with BMI >25 had larger tumors, more positive lymph nodes, higher grade breast cancer, higher ki67 which indicates more aggressive growth and characteristics of cancer cells. No conflict of interest. 1897 POSTER RAD001, A mTOR specificity inhibitor, combined with PI3K Inhibitor LY294002, significantly reduced proliferation and induced apoptosis of triple negative breast cancer cell lines MDA-MB-231 through PI3K/Akt/mTOR pathway in vitro G. Sun1,2 , B. Ma1 , L. Yang1 , C. Dong1 , B. Ma1,2 . 1 The Affiliated Tumor Hospital of Xinjing Medical University, Breast and Head-neck Oncology, Urumqi, China; 2 Xinjiang Institute of Cancer Prevention and Treatment, Breast Cancer Institute, Urumqi, China Background: Research of targeted therapies about PI3K/Akt/mTOR pathway have became a hotspot recently, and related researches have been extended from a single-target suppression to multiple-targets combined inhibition. In this study, by using LY294002 and RAD001, the specific inhibitors of PI3K and mTOR, to observe whether the anti-tumor effects on the proliferation, cycle distribution and apoptosis of combined inhibitors were a synergistic effect, and were differences between different molecular characteristics of human breast cancer cells alone and in combination in vitro. Materials and Methods: Routinely cultured MCF-7, SK-BR-3 and MDAMB-231 cells in vitro. Logarithmic phase cells among each cells were selected and divided into the blank control group, LY294002 group, RAD001 group and the combination group. MTT assay and flow cytometry were used to detect the cell proliferation, cell cycle distribution and cell apoptosis of different groups. Results: (1) LY294002 and RAD001 could significantly inhibit the proliferations of MCF-7, SK-BR-3 and MDA-MB-231 cells with a dosedependent manner respectively (P < 0.05), and compared with other cells, MDA-MB-231 cells were more sensitive to both drugs (P < 0.05). The antitumor effects were significantly increased in combination groups of different cells, and showed an additive effects. (2) MCF-7, SK-BR-3 and MDA-MB231 cells could be arrest in G1 phase by IC50 of LY294002 and RAD001 respectively(P < 0.05), and the effects of combination groups were more significantly compared with monotherapy groups(P < 0.05). However, no significant differences among different cells. (3) LY294002 and RAD001 could significantly increase the apoptosis rates of human MCF-7, SK-BR-3 and MDA-MB-231 cells alone (P < 0.05), but there were no differences between different cells. When two inhibitors were combined, the apoptosis rates of the different cells were significantly increased compared with single drug, especially in MDA-MB-231 cell. The apoptosis rates of MCF-7, SK-BR-3 and MDA-MB-231 cells were 17.58%, 44.28% and 52.67% respectively, and there were significantly different between different cell lines (P < 0.05). Conclusions: Targeted therapy on P13K/Akt/mTOR pathway could significantly inhibit the proliferation of human breast cancer cell lines by inducing apoptosis and arresting cell cycle distribution. By using a combination of different inhibitors which were targeted on different related genes of this pathway such as PI3K and mTOR, the anti-tumor effects were more significant increasd compared with monotherapy, especially in MDA-MB-231 cell line which were negative with ER, PR and HER-2. It was important to provide new research ideas for individualized treatment and translational medicine of breast cancer. No conflict of interest. 1898 POSTER Characteristics of the local immunity of breast cancer patients with different reproductive function statuses Y. Shatova1 , E. Zlatnik2 , I. Novikova2 , A. Bakhtin2 . 1 Rostov Research Institute of Oncology, Soft Tissue Department, Rostov-Don, Russian Federation; 2 Rostov Research Institute of Oncology, Department of Immunology, Rostov-Don, Russian Federation Background: Local immunity factors play an important role in biological behavior of tumor, and this role depends on a patient’s state. Ovarianmenstrual function determined by hormonal profile is clinically significant for breast cancer (BC) development. The purpose of the study was to assess some parameters of the local cellular immunity in BC in patients with different ovarian-menstrual function statuses. Material and Methods: Tumor tissue samples obtained during the surgery from 95 patients (15 with maintained function, 27 perimenopausal, 53
Abstracts postmenopausal) were used as material for the study. Populations and subpopulations of lymphocytes were determined in tissue homogenates using FACSCantoII (BD) flow cytometer. Of the total number of CD45+ lymphocytes, the percentage of lymphocytes expressing CD3, CD4, CD8, CD19, CD16/56 markers was calculated. Results: There were no statistically significant differences in CD3+ levels in tumor tissues in dependence on the reproductive status of the patients, though T-lymphocyte subpopulations differed in the reproductive and menopausal groups. A higher content of CD3+CD8+ was observed in tumors of patients of the reproductive group in comparison with the perimenopausal and postmenopausal groups (56.5±7.3, 37.2±2.95 and 37.4±2.1%, respectively; p < 0.05). Level of CD3+CD4+ cells was lower in patients with maintained ovarian-menstrual function (34.6±6.4% against 50.3±3.2% in perimenopausal and 45.4±1.55% in postmenopausal patients). The postmenopausal group was characterized by a lower content of NK cells in tumor in comparison with the reproductive group, except for luminal A breast cancer in which levels of NK cells did not differ in postmenopausal and reproductive patients. The other differences were observed in all biological subtypes of BC (luminal A, luminal B, HER2-neu+ and triple negative BC). Conclusions: Lymphocytic content in BC tissue depends on the reproductive status of patients. No conflict of interest. 1899 POSTER Correlation between Ki-67 and FISH testing of HER2 IHC 1+ early invasive breast cancer A.C. Latorre1 , L. Caldarola2 , S. Petroni2 , F. Giotta1 , V. Lorusso1 , I. Trotti2 , G. Simone2 . 1 IRCCS Onc. Pap Giovanni Paolo II, Medical Oncology Unit, Bari, Italy; 2 IRCCS Onc. Pap Giovanni Paolo II, Pathology Department, Bari, Italy. Background: HER2 gene amplification or overexpression in invasive breast cancer (IBC) has been demonstrated to be a parameter for bad prognosis. Diagnostic assays for HER2 expression have a high predictive value because HER2 positive tumors can benefit from target therapy with trastuzumab.The aim of the present study is to analyze the incidence of HER2 gene amplification and to assess if it would be clinically useful to test HER2 gene amplification in selected tumors with adverse prognostic features scoring 1+ by immunohistochemistry (IHC). Material and Methods: Seventy-five cases of early IBC in women who underwent elective surgery in 2013 entered the study. According to the histotype, 61 tumors were classified as infiltrating ductal carcinoma (IDC) and 14 as infiltrating lobular carcinoma (ILC) and received a HER2 score of 1+ by IHC. Among these, 48 IBC with unfavorable prognostic tumor characteristics were selected and tested by FISH. HER2 amplification was evaluated using the Vysis HER2/Cep17 probe (Path Vysion HER2 DNA Probe Kit® , Abbott Molecular, IL). In agreement with the ASCO/CAP/SIAPEC guidelines, HER2 ratio–based amplification was considered. Gene amplification was evaluated as present when the HER2/Cep17 ratio was 2 or more or when the mean HER2 copy number was more than 6. Results: In 2013, 331 consecutive IBC were tested by IHC for HER2 and 102 cases were scored (31%) 1+. We selected 75 patients, 61 IDC and 14 ILC (81% and 19% respectively), out of 102 cases scoring 1+ who had undergone radical elective surgery. Forty-eight IBC samples out of 75 (64%) (42 IDC and 6 ILC) were selected according to one or more unfavorable prognostic tumor characteristics (histological grade, proliferative index, absent hormone receptor expression, node positivity and vascular invasion); 22 (46%) showed high histological grade (G3), 23 (48%) had a high proliferative index (Ki-6730%), 32 (67%) were nodepositive, 27 (56%) showed vascular invasion; regarding hormone receptors expression, 3 (6%) showed no ER expression and 10 (21%) showed no PgR expression. FISH was performed on 48 IBC scoring 1+ by IHC and 7 IDC out of 48 (14.6%) showed HER2 amplification; all 7 samples showed a high proliferative index. In this subgroup, the statistical analysis with Fisher’s exact test evidenced a significant association between the presence of gene amplification and high proliferative index (P = 0.0094). Conclusions: Our retrospective data suggest that early IDC patients scoring HER2 1+ by IHC who show Ki-6730% must be tested by FISH because there is a significant association between HER2 amplification and high proliferative index. In this subgroup of patients Ki-6730% would represent a predictive factor of HER2 amplification. No conflict of interest.
S299 1900 POSTER Real-time RT-PCR quantification of human telomerase reverse transcriptase in breast cancer H. Abdel Raouf Shaban1 , K. Amr2 , M. Afify3 . 1 National Research Center, Immunogenetics, Giza, Egypt; 2 National Research Center, Medical Molecular Genetics, Giza, Egypt; 3 National Research Center, Biochemistry, Giza, Egypt Background: Telomerase is a cellular ribonucleoprotein reverse transcriptase (TERT), and telomerase RNA segments act as templates for the synthesis of telomeric DNA onto chromosomal ends. The enzyme telomerase catalyzes the de novo synthesis of telomere repeats, thereby maintaining telomere length, which is necessary for unlimited cellular proliferation. Human TERT is often detectable in cancerous cells but not in normal somatic cells Thus, hTERT has been proposed as a marker with diagnostic and prognostic potential in breast cancer. This study aimed to investigate expression of hTERT mRNA in peripheral blood and tissue of breast cancer patients and to present strategies for early detection screen test. Methods: Twenty-two breast cancer patients and twelve healthy individuals were studied. Expression of hTERT mRNA and GAPDH (RNA, as reference gene) were analyzed using quantitative real-time reverse transcriptasepolymerase chain reaction (RT-PCR) on total RNA from blood and tissue of breast cancer patients. An expression index (EI) was calculated for telomerase expression. Results: According to RT-qPCR results, the mean values of hTERT expression in the blood and tissue samples of the breast cancer patient were (5.74 EI & 59.66 EI) respectively where in healthy donors was (1.87 EI). The results also demonstrated that, hTERT expression in breast cancer patient showed 3.07 fold increases in relation to control. Conclusion: These results indicated that, telomerase activity could be a promising tumor marker for breast cancer. Also, the present results suggest the feasibility of using RT-PCR in routine screening of telomerase activity in blood specimens. However, more investigations with larger numbers of samples are needed to verify these results. No conflict of interest. 1901 POSTER Mammary tuberculosis in females: Simulating malignant lesion of the breast A. Setia1 , Sciences, Sciences, Sciences,
S.B. Sharma2 , G. Nirwal3 . 1 National institute of Medial General Surgery, Jaipur, India; 2 National Institute of Medical General Surgery, Jaipur, India; 3 National Institute of Medical General Seurgery, Jaipur, India
Background: Mammary tuberculosis is a rare entity and may be mistaken for malignant lesion of the breast. Our study analyses the clinical presentation of mammary tuberculosis mimicking malignancy of breast and the methodical diagnostic modalities in 5 such patients. Materials and Methods: In the year 2012–2014, five patients of Mammary Tuberculosis were treated at NIMS medical college, Jaipur, India and were included in this study. All the patients underwent an elaborate clinical examination, blood investigations, FNAC (Fine needle aspiration cytology) and/or Core biopsy and Ultrasonography. Results: Four patients presented with a lump, one of these had secondary skin changes, and fifth patient presented with diffuse swelling of the entire breast (n = 4+1). Associated pain was present in two patients and constitutional symptoms in three and two patients gave history of current or recent Lactation. Demonstration of Acid fast bacilli on staining and/or giant cells, Epithelioid granuloma and/or necrosis on histopathology clenched the diagnosis. Anti-tubercular therapy initiated after confirmation was the mainstay of treatment and resulted in complete resolution of the lump. Conclusion: Mammary Tuberculosis can mimic malignancy of breast clouding the clinical diagnosis. A step-wise approach in the form of clinical examination, radiological studies and histopathology and staining should be followed. USG is usually inconclusive but is very useful in obtaining core biopsy tissue samples. Unnecessary open biopsies should be avoided as far as possible. Anti-tubercular therapy should be initiated immediately after confirmation of the diagnosis. Surgical interventions can also be resorted to in non-responsive cases. No conflict of interest.
Poster Session (Monday, 28 September) Breast Cancer − Early Disease 1896 POSTER Correlation between body mass index (BMI) and histopathologic characteristics of breast cancer and recurrence rate in pre and postmenopausal women W. El-Sadda1 , I. Abdel-Halim2 , M. Magdy2 , M. El-Ibrashy2 , H. Daghriri1 . 1 Al-Ghad International Colleges for Health Sciences, Radiology, Najran, Saudi Arabia; 2 Mansoura University Hospital, Clinical Oncology & Nuclear Medicine, Mansoura, Egypt Background: High BMI is related to increased breast cancer incidence to more aggressive and poorer prognosis, The study aims to analyze the association between increased body weight by using the body mass index (BMI) at time of diagnosis and histopathologic features of breast cancer (tumor size, nuclear grade, hormone receptors (ER & PR), HER-2/neu expression, Ki-67 index, lymphatic/vascular invasion, axillary nodes involvement and menopausal status, to evaluate the impact of BMI on DFS at multivariate analysis. Patients and Methods: In this retrospective study, 500 patients with early breast cancer (200 premenopausal and 300 postmenopausal) were classified into subgroups according to BMI; normal was defined as BMI <25 kg/m2 , overweight as BMI >25 kg/m2 and obese as BMI >30 kg/m2 . The association between BMI and histopathologic features, ki-67 labeling index was evaluated. Results: High BMI significantly associated with larger size tumor both in premenopausal (p = 0.01) and postmenopausal women (p = 0.001) Obese premenopausal women were associated with more aggressive histopathologic characteristics (e.g., more positive axillary nodes p = 0.016 and presence of vascular invasion p = 0.005) compared to normal weight group. Postmenopausal patients with BMI >25 developed more frequently ER/PR positive cancer (85% vs. 70%, p = 0.01) while no association was found in premenopausal women. BMI did not show significant association to tumor nuclear grade, HER-/neu overexpression, Ki-67 index and lymphatic invasion in both pre and postmenopausal women. In multivariate analysis, obesity was not an independent prognostic factor in pre and postmenopause, however, obese women showed a shorter DFS and in the postmenopausal group with BMI >25 as a single variable had a statistically significant worse DFS compared to normal weight women.
Abstracts Conclusion: Obesity plays a different role in breast cancer according to menopausal status; postmenopausal women with BMI >25 had larger tumors, more positive lymph nodes, higher grade breast cancer, higher ki67 which indicates more aggressive growth and characteristics of cancer cells. No conflict of interest. 1897 POSTER RAD001, A mTOR specificity inhibitor, combined with PI3K Inhibitor LY294002, significantly reduced proliferation and induced apoptosis of triple negative breast cancer cell lines MDA-MB-231 through PI3K/Akt/mTOR pathway in vitro G. Sun1,2 , B. Ma1 , L. Yang1 , C. Dong1 , B. Ma1,2 . 1 The Affiliated Tumor Hospital of Xinjing Medical University, Breast and Head-neck Oncology, Urumqi, China; 2 Xinjiang Institute of Cancer Prevention and Treatment, Breast Cancer Institute, Urumqi, China Background: Research of targeted therapies about PI3K/Akt/mTOR pathway have became a hotspot recently, and related researches have been extended from a single-target suppression to multiple-targets combined inhibition. In this study, by using LY294002 and RAD001, the specific inhibitors of PI3K and mTOR, to observe whether the anti-tumor effects on the proliferation, cycle distribution and apoptosis of combined inhibitors were a synergistic effect, and were differences between different molecular characteristics of human breast cancer cells alone and in combination in vitro. Materials and Methods: Routinely cultured MCF-7, SK-BR-3 and MDAMB-231 cells in vitro. Logarithmic phase cells among each cells were selected and divided into the blank control group, LY294002 group, RAD001 group and the combination group. MTT assay and flow cytometry were used to detect the cell proliferation, cell cycle distribution and cell apoptosis of different groups. Results: (1) LY294002 and RAD001 could significantly inhibit the proliferations of MCF-7, SK-BR-3 and MDA-MB-231 cells with a dosedependent manner respectively (P < 0.05), and compared with other cells, MDA-MB-231 cells were more sensitive to both drugs (P < 0.05). The antitumor effects were significantly increased in combination groups of different cells, and showed an additive effects. (2) MCF-7, SK-BR-3 and MDA-MB231 cells could be arrest in G1 phase by IC50 of LY294002 and RAD001 respectively(P < 0.05), and the effects of combination groups were more significantly compared with monotherapy groups(P < 0.05). However, no significant differences among different cells. (3) LY294002 and RAD001 could significantly increase the apoptosis rates of human MCF-7, SK-BR-3 and MDA-MB-231 cells alone (P < 0.05), but there were no differences between different cells. When two inhibitors were combined, the apoptosis rates of the different cells were significantly increased compared with single drug, especially in MDA-MB-231 cell. The apoptosis rates of MCF-7, SK-BR-3 and MDA-MB-231 cells were 17.58%, 44.28% and 52.67% respectively, and there were significantly different between different cell lines (P < 0.05). Conclusions: Targeted therapy on P13K/Akt/mTOR pathway could significantly inhibit the proliferation of human breast cancer cell lines by inducing apoptosis and arresting cell cycle distribution. By using a combination of different inhibitors which were targeted on different related genes of this pathway such as PI3K and mTOR, the anti-tumor effects were more significant increasd compared with monotherapy, especially in MDA-MB-231 cell line which were negative with ER, PR and HER-2. It was important to provide new research ideas for individualized treatment and translational medicine of breast cancer. No conflict of interest. 1898 POSTER Characteristics of the local immunity of breast cancer patients with different reproductive function statuses Y. Shatova1 , E. Zlatnik2 , I. Novikova2 , A. Bakhtin2 . 1 Rostov Research Institute of Oncology, Soft Tissue Department, Rostov-Don, Russian Federation; 2 Rostov Research Institute of Oncology, Department of Immunology, Rostov-Don, Russian Federation Background: Local immunity factors play an important role in biological behavior of tumor, and this role depends on a patient’s state. Ovarianmenstrual function determined by hormonal profile is clinically significant for breast cancer (BC) development. The purpose of the study was to assess some parameters of the local cellular immunity in BC in patients with different ovarian-menstrual function statuses. Material and Methods: Tumor tissue samples obtained during the surgery from 95 patients (15 with maintained function, 27 perimenopausal, 53
Abstracts postmenopausal) were used as material for the study. Populations and subpopulations of lymphocytes were determined in tissue homogenates using FACSCantoII (BD) flow cytometer. Of the total number of CD45+ lymphocytes, the percentage of lymphocytes expressing CD3, CD4, CD8, CD19, CD16/56 markers was calculated. Results: There were no statistically significant differences in CD3+ levels in tumor tissues in dependence on the reproductive status of the patients, though T-lymphocyte subpopulations differed in the reproductive and menopausal groups. A higher content of CD3+CD8+ was observed in tumors of patients of the reproductive group in comparison with the perimenopausal and postmenopausal groups (56.5±7.3, 37.2±2.95 and 37.4±2.1%, respectively; p < 0.05). Level of CD3+CD4+ cells was lower in patients with maintained ovarian-menstrual function (34.6±6.4% against 50.3±3.2% in perimenopausal and 45.4±1.55% in postmenopausal patients). The postmenopausal group was characterized by a lower content of NK cells in tumor in comparison with the reproductive group, except for luminal A breast cancer in which levels of NK cells did not differ in postmenopausal and reproductive patients. The other differences were observed in all biological subtypes of BC (luminal A, luminal B, HER2-neu+ and triple negative BC). Conclusions: Lymphocytic content in BC tissue depends on the reproductive status of patients. No conflict of interest. 1899 POSTER Correlation between Ki-67 and FISH testing of HER2 IHC 1+ early invasive breast cancer A.C. Latorre1 , L. Caldarola2 , S. Petroni2 , F. Giotta1 , V. Lorusso1 , I. Trotti2 , G. Simone2 . 1 IRCCS Onc. Pap Giovanni Paolo II, Medical Oncology Unit, Bari, Italy; 2 IRCCS Onc. Pap Giovanni Paolo II, Pathology Department, Bari, Italy. Background: HER2 gene amplification or overexpression in invasive breast cancer (IBC) has been demonstrated to be a parameter for bad prognosis. Diagnostic assays for HER2 expression have a high predictive value because HER2 positive tumors can benefit from target therapy with trastuzumab.The aim of the present study is to analyze the incidence of HER2 gene amplification and to assess if it would be clinically useful to test HER2 gene amplification in selected tumors with adverse prognostic features scoring 1+ by immunohistochemistry (IHC). Material and Methods: Seventy-five cases of early IBC in women who underwent elective surgery in 2013 entered the study. According to the histotype, 61 tumors were classified as infiltrating ductal carcinoma (IDC) and 14 as infiltrating lobular carcinoma (ILC) and received a HER2 score of 1+ by IHC. Among these, 48 IBC with unfavorable prognostic tumor characteristics were selected and tested by FISH. HER2 amplification was evaluated using the Vysis HER2/Cep17 probe (Path Vysion HER2 DNA Probe Kit® , Abbott Molecular, IL). In agreement with the ASCO/CAP/SIAPEC guidelines, HER2 ratio–based amplification was considered. Gene amplification was evaluated as present when the HER2/Cep17 ratio was 2 or more or when the mean HER2 copy number was more than 6. Results: In 2013, 331 consecutive IBC were tested by IHC for HER2 and 102 cases were scored (31%) 1+. We selected 75 patients, 61 IDC and 14 ILC (81% and 19% respectively), out of 102 cases scoring 1+ who had undergone radical elective surgery. Forty-eight IBC samples out of 75 (64%) (42 IDC and 6 ILC) were selected according to one or more unfavorable prognostic tumor characteristics (histological grade, proliferative index, absent hormone receptor expression, node positivity and vascular invasion); 22 (46%) showed high histological grade (G3), 23 (48%) had a high proliferative index (Ki-6730%), 32 (67%) were nodepositive, 27 (56%) showed vascular invasion; regarding hormone receptors expression, 3 (6%) showed no ER expression and 10 (21%) showed no PgR expression. FISH was performed on 48 IBC scoring 1+ by IHC and 7 IDC out of 48 (14.6%) showed HER2 amplification; all 7 samples showed a high proliferative index. In this subgroup, the statistical analysis with Fisher’s exact test evidenced a significant association between the presence of gene amplification and high proliferative index (P = 0.0094). Conclusions: Our retrospective data suggest that early IDC patients scoring HER2 1+ by IHC who show Ki-6730% must be tested by FISH because there is a significant association between HER2 amplification and high proliferative index. In this subgroup of patients Ki-6730% would represent a predictive factor of HER2 amplification. No conflict of interest.
S299 1900 POSTER Real-time RT-PCR quantification of human telomerase reverse transcriptase in breast cancer H. Abdel Raouf Shaban1 , K. Amr2 , M. Afify3 . 1 National Research Center, Immunogenetics, Giza, Egypt; 2 National Research Center, Medical Molecular Genetics, Giza, Egypt; 3 National Research Center, Biochemistry, Giza, Egypt Background: Telomerase is a cellular ribonucleoprotein reverse transcriptase (TERT), and telomerase RNA segments act as templates for the synthesis of telomeric DNA onto chromosomal ends. The enzyme telomerase catalyzes the de novo synthesis of telomere repeats, thereby maintaining telomere length, which is necessary for unlimited cellular proliferation. Human TERT is often detectable in cancerous cells but not in normal somatic cells Thus, hTERT has been proposed as a marker with diagnostic and prognostic potential in breast cancer. This study aimed to investigate expression of hTERT mRNA in peripheral blood and tissue of breast cancer patients and to present strategies for early detection screen test. Methods: Twenty-two breast cancer patients and twelve healthy individuals were studied. Expression of hTERT mRNA and GAPDH (RNA, as reference gene) were analyzed using quantitative real-time reverse transcriptasepolymerase chain reaction (RT-PCR) on total RNA from blood and tissue of breast cancer patients. An expression index (EI) was calculated for telomerase expression. Results: According to RT-qPCR results, the mean values of hTERT expression in the blood and tissue samples of the breast cancer patient were (5.74 EI & 59.66 EI) respectively where in healthy donors was (1.87 EI). The results also demonstrated that, hTERT expression in breast cancer patient showed 3.07 fold increases in relation to control. Conclusion: These results indicated that, telomerase activity could be a promising tumor marker for breast cancer. Also, the present results suggest the feasibility of using RT-PCR in routine screening of telomerase activity in blood specimens. However, more investigations with larger numbers of samples are needed to verify these results. No conflict of interest. 1901 POSTER Mammary tuberculosis in females: Simulating malignant lesion of the breast A. Setia1 , Sciences, Sciences, Sciences,
S.B. Sharma2 , G. Nirwal3 . 1 National institute of Medial General Surgery, Jaipur, India; 2 National Institute of Medical General Surgery, Jaipur, India; 3 National Institute of Medical General Seurgery, Jaipur, India
Background: Mammary tuberculosis is a rare entity and may be mistaken for malignant lesion of the breast. Our study analyses the clinical presentation of mammary tuberculosis mimicking malignancy of breast and the methodical diagnostic modalities in 5 such patients. Materials and Methods: In the year 2012–2014, five patients of Mammary Tuberculosis were treated at NIMS medical college, Jaipur, India and were included in this study. All the patients underwent an elaborate clinical examination, blood investigations, FNAC (Fine needle aspiration cytology) and/or Core biopsy and Ultrasonography. Results: Four patients presented with a lump, one of these had secondary skin changes, and fifth patient presented with diffuse swelling of the entire breast (n = 4+1). Associated pain was present in two patients and constitutional symptoms in three and two patients gave history of current or recent Lactation. Demonstration of Acid fast bacilli on staining and/or giant cells, Epithelioid granuloma and/or necrosis on histopathology clenched the diagnosis. Anti-tubercular therapy initiated after confirmation was the mainstay of treatment and resulted in complete resolution of the lump. Conclusion: Mammary Tuberculosis can mimic malignancy of breast clouding the clinical diagnosis. A step-wise approach in the form of clinical examination, radiological studies and histopathology and staining should be followed. USG is usually inconclusive but is very useful in obtaining core biopsy tissue samples. Unnecessary open biopsies should be avoided as far as possible. Anti-tubercular therapy should be initiated immediately after confirmation of the diagnosis. Surgical interventions can also be resorted to in non-responsive cases. No conflict of interest.