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Lymphocyte subpopulations in peripheral blood of breast cancer patients
Abstracts / The Breast 20 (2011) S12–S55
Results: Similarly, association between p53 protein and disease response was evaluated. With 90% confidence it was established, that p53 protein overexpression influences disease progression (Fisher‘s p ¼ 0.096). We also estimated the impact of protein expression on disease outcomes in 24 patients who underwent only first-line treatment against metastatic breast cancer. If p53 protein overexpression was found, mean survival was 28.5 months (SEM ¼ 5.4, 95% CI 17.9–39.0) and median 25 months (SEM ¼ 6.9, 95% CI 11.4–38.6), if it was absent – survival was slightly longer: mean 33.6 months (SEM ¼ 4.2, 95% CI 25.3–41.8) and median 30 months (SEM ¼ 2.6, 95% CI 24.9– 35.0). Kaplan-Meier method did not detect statistically significant differences between the groups (log-rank p > 0.05, Breslow p > 0.05). For small samples, associations among molecular markers were evaluated using Fisher‘s exact test. It was established, that p53 protein has a trend to be associated with underexpression of ER (OR ¼ 5.83, 95% CI 0.9–37.81, p ¼ 0.064). Associations among other factors were non-significant. Analysis of combinations of ER and PR (ER-PR-, ER-PR+, ER+PR-, ER+PR+) with p53 protein and HER2 was also conducted. However, there were no significant association between them. Additionally, combination of negative expression in 3 markers (ER, PR and HER2) with p53 protein expression was checked. Thus, significant association was found between positive p53 protein expression and combination of molecular markers ER-PRHER2- (Fisher‘s p ¼ 0.041). Conclusion: It was established, that combination of molecular markers ER-PR-HER2- is 7.7 times (95% CI 1.16–51.17; p ¼ 0.035) more likely in case of positive expression of p53 protein and it is associated with the worse prognosis of the patients.
PO38 LYMPHOCYTE SUBPOPULATIONS IN PERIPHERAL BLOOD OF BREAST CANCER PATIENTS Lidia Skotarenko, Zaira Kadagidze, Olga Korotkova, Igor Vorotnikov, Nino Chkhikvadze, Maria Rodionova N.N.Blokhin Russian Cancer Center, Moscow, Russia, Moscow, Russia Purpose: To study structure of lymphocytes subpopulation of breast cancer patients. Materials and methods: Studied linear markers of cellular immunity and estimated subpopulation structure of peripheral blood lymphocytes of patients with breast cancer, using multiparameter analysis of immunophenotype of lymphocytes. The study included 77 patients with Stage I (T1N0) - 31,2%, and Stage II (T1N1, T2N0 – N1) - 19,5%. Results: The study revealed that 58,4% of patients are violations of the structure of T lymphocytes compared with healthy donors. Surgical treatment had no effect on the dynamics of the distribution of major lymphocyte populations in the analysis of the total group of patients. Individual analysis of immunograms depending on the initial level of CD3 showed that 28,5% of patients were initially reduced the content of CD3+, at 41,5% - initially normal number of CD3+ lymphocytes, at 30,0% - initially higher amount. In patients with reduced CD3+ lymphocytes was initially increases the number of NK cells with the phenotype CD3-CD16+CD56+, whereas in patients with normal and high numbers of CD3+ lymphocytes was significantly increased the number of NK cells with phenotype CD3+CD16+CD56+, 14,01,1 and 19,31,4, respectively. Conclusion: To provide adequate chemo- and radiotherapy, monitoring of immune system is to be held for further correction of subpopulation structure of T cell immunity.
PO39 LOSS OF TRAP1 EXPRESSION AS A POSSIBLE MECHANISM BEHIND RECURRENCE RISK IN ADVANCED BREAST CANCER Ern Yu Tan 1, Francesco Pezzella 2, Kevin Gatter 2 Tan Tock Seng Hospital, Singapore, Singapore 2 Nuffield Department of Clinical Laboratory Sciences, University of Oxford, Oxford, UK 1
S25
Introduction: Tumours with N2 nodal involvement have a 3-fold increase in recurrence compared to those with N1 or N0 disease. Because most disease recurrences in N2 disease occur in distant sites, recurrence signifies a poor outcome. The Tumour Necrosis Factor Receptor Associated Protein 1 (TRAP1) has been proposed to have a novel role in breast cancer. TRAP1 is a molecular chaperone of the tumour suppressor Retinoblastoma (Rb) protein and plays a role in Rb regulation of G1/S transition. Loss of TRAP1 function may therefore represent a novel oncogenic pathway where the loss of function of a tumour suppressor gene occurs through the loss of its chaperone. We therefore aim to evaluate the role of TRAP1 in breast tumourogenesis and progression. Materials and Methods: TRAP1 and Rb expression were evaluated in a well-characterised series of invasive breast cancers with N2 nodal involvement using immumohistochemistry. TRAP1 levels were evaluated in 7 breast cancer cell lines. Interaction between Rb and E2F1 and changes in S phase fraction on flow cytometry were examined following TRAP1 silencing and over-expression. The effect of TRAP1 expression on cellular invasion was examined in invasive assays. Results: TRAP1 was expressed in the nucleus in 54% of tumours, and in the cytoplasm of 65%, where a granular pattern suggestive of mitochondrial localisation was observed. Nulcear TRAP1 expression was found to correlate with Rb expression (P <0.01), consistent with its role as a molecular chaperone of Rb. Loss of nuclear TRAP1 expression was associated with an increased risk of recurrence and a shorter disease-free survival, an effect that was particularly seen in the first 5 years (P ¼ 0.04). TRAP1 protein and mRNA levels were evaluated in 7 breast cancer cell lines. TRAP1 levels were significantly lower in the MDA468 and MDA231 cell lines, both of which were hormone unresponsive. In hypoxia, TRAP1 was observed to translocate into the nucleus where it associated with Rb. TRAP1 silencing attenuated Rb-E2F1 interaction, resulting in a larger fraction of cells entering into the S phase. Conversely, when TRAP1 was re-expressed in TRAP1-deficient MDA231 breast cancer cells, Rb-E2F1 interaction was enhanced and a smaller S-phase fraction was observed. TRAP1 also appears important in cellular invasion. The invasiveness of MDA231 cells appeared to be attenuated by the restoration of TRAP1, and was possibly due to a reduction in MMP9 levels. Conclusions: TRAP1 appears to modulate tumour behaviour in response to hypoxic stress. Loss of TRAP1 expression increased the proliferative and invasive potential of breast cancer cells, and was associated with a poorer disease-free survival in N2 disease particularly within the first 5 years.
PO40 KI-67 EXPRESSION PREDICTS ADVANCED BREAST CANCER
CLINICAL
OUTCOME
IN
LOCALLY
Naomi Kobayashi, Masahiro Hikichi, Shinsuke Miyajima, Kaori Ushimado, Toshiaki Utsumi Fujita Health University, Dept. of Breast Surgery, Toyoake, Aichi, Japan Goals: The management of locally advanced breast cancer (LABC) requires a combined treatment approach involving surgery, radiation therapy and systemic therapy. Recently, the Ki-67 expression has been considered as an important factor for selecting the additional chemotherapy to endocrine therapy in hormone receptor positive breast cancer. The aim of this study was to examine the prognostic significance of the Ki-67 in LABC. Materials and Methods: One thousand six hundred and twenty patients were diagnosed with breast cancer at our institution from 1992 to 2010. We conducted a retrospective analysis of the 140 patients with LABC (Stage IIIA, IIIB, and IIIC) in this period. The median age of the patients was 56 years with a range of 22 to 92 years old. The expression of Ki-67 was assessed by immunohistochemistry with a monoclonal MIB1 antibody. Ki67 labeling index (LI) was categorized as low (<15%) and high (> or ¼15%) in LABC. Disease-free survival (DFS) curves and over-all survival (OS) curves were estimated using the Kaplan-Meier method and survival comparisons were made with the log-rank test. A multivariate Cox proportional hazards regression model performed in a stepwise fashion was used to determine the prognostic value of each significant variable. The level of significance was taken to be 0.05. IBM SPSS Statistics 19 software package was used for statistical analysis.
Results: Similarly, association between p53 protein and disease response was evaluated. With 90% confidence it was established, that p53 protein overexpression influences disease progression (Fisher‘s p ¼ 0.096). We also estimated the impact of protein expression on disease outcomes in 24 patients who underwent only first-line treatment against metastatic breast cancer. If p53 protein overexpression was found, mean survival was 28.5 months (SEM ¼ 5.4, 95% CI 17.9–39.0) and median 25 months (SEM ¼ 6.9, 95% CI 11.4–38.6), if it was absent – survival was slightly longer: mean 33.6 months (SEM ¼ 4.2, 95% CI 25.3–41.8) and median 30 months (SEM ¼ 2.6, 95% CI 24.9– 35.0). Kaplan-Meier method did not detect statistically significant differences between the groups (log-rank p > 0.05, Breslow p > 0.05). For small samples, associations among molecular markers were evaluated using Fisher‘s exact test. It was established, that p53 protein has a trend to be associated with underexpression of ER (OR ¼ 5.83, 95% CI 0.9–37.81, p ¼ 0.064). Associations among other factors were non-significant. Analysis of combinations of ER and PR (ER-PR-, ER-PR+, ER+PR-, ER+PR+) with p53 protein and HER2 was also conducted. However, there were no significant association between them. Additionally, combination of negative expression in 3 markers (ER, PR and HER2) with p53 protein expression was checked. Thus, significant association was found between positive p53 protein expression and combination of molecular markers ER-PRHER2- (Fisher‘s p ¼ 0.041). Conclusion: It was established, that combination of molecular markers ER-PR-HER2- is 7.7 times (95% CI 1.16–51.17; p ¼ 0.035) more likely in case of positive expression of p53 protein and it is associated with the worse prognosis of the patients.
PO38 LYMPHOCYTE SUBPOPULATIONS IN PERIPHERAL BLOOD OF BREAST CANCER PATIENTS Lidia Skotarenko, Zaira Kadagidze, Olga Korotkova, Igor Vorotnikov, Nino Chkhikvadze, Maria Rodionova N.N.Blokhin Russian Cancer Center, Moscow, Russia, Moscow, Russia Purpose: To study structure of lymphocytes subpopulation of breast cancer patients. Materials and methods: Studied linear markers of cellular immunity and estimated subpopulation structure of peripheral blood lymphocytes of patients with breast cancer, using multiparameter analysis of immunophenotype of lymphocytes. The study included 77 patients with Stage I (T1N0) - 31,2%, and Stage II (T1N1, T2N0 – N1) - 19,5%. Results: The study revealed that 58,4% of patients are violations of the structure of T lymphocytes compared with healthy donors. Surgical treatment had no effect on the dynamics of the distribution of major lymphocyte populations in the analysis of the total group of patients. Individual analysis of immunograms depending on the initial level of CD3 showed that 28,5% of patients were initially reduced the content of CD3+, at 41,5% - initially normal number of CD3+ lymphocytes, at 30,0% - initially higher amount. In patients with reduced CD3+ lymphocytes was initially increases the number of NK cells with the phenotype CD3-CD16+CD56+, whereas in patients with normal and high numbers of CD3+ lymphocytes was significantly increased the number of NK cells with phenotype CD3+CD16+CD56+, 14,01,1 and 19,31,4, respectively. Conclusion: To provide adequate chemo- and radiotherapy, monitoring of immune system is to be held for further correction of subpopulation structure of T cell immunity.
PO39 LOSS OF TRAP1 EXPRESSION AS A POSSIBLE MECHANISM BEHIND RECURRENCE RISK IN ADVANCED BREAST CANCER Ern Yu Tan 1, Francesco Pezzella 2, Kevin Gatter 2 Tan Tock Seng Hospital, Singapore, Singapore 2 Nuffield Department of Clinical Laboratory Sciences, University of Oxford, Oxford, UK 1
S25
Introduction: Tumours with N2 nodal involvement have a 3-fold increase in recurrence compared to those with N1 or N0 disease. Because most disease recurrences in N2 disease occur in distant sites, recurrence signifies a poor outcome. The Tumour Necrosis Factor Receptor Associated Protein 1 (TRAP1) has been proposed to have a novel role in breast cancer. TRAP1 is a molecular chaperone of the tumour suppressor Retinoblastoma (Rb) protein and plays a role in Rb regulation of G1/S transition. Loss of TRAP1 function may therefore represent a novel oncogenic pathway where the loss of function of a tumour suppressor gene occurs through the loss of its chaperone. We therefore aim to evaluate the role of TRAP1 in breast tumourogenesis and progression. Materials and Methods: TRAP1 and Rb expression were evaluated in a well-characterised series of invasive breast cancers with N2 nodal involvement using immumohistochemistry. TRAP1 levels were evaluated in 7 breast cancer cell lines. Interaction between Rb and E2F1 and changes in S phase fraction on flow cytometry were examined following TRAP1 silencing and over-expression. The effect of TRAP1 expression on cellular invasion was examined in invasive assays. Results: TRAP1 was expressed in the nucleus in 54% of tumours, and in the cytoplasm of 65%, where a granular pattern suggestive of mitochondrial localisation was observed. Nulcear TRAP1 expression was found to correlate with Rb expression (P <0.01), consistent with its role as a molecular chaperone of Rb. Loss of nuclear TRAP1 expression was associated with an increased risk of recurrence and a shorter disease-free survival, an effect that was particularly seen in the first 5 years (P ¼ 0.04). TRAP1 protein and mRNA levels were evaluated in 7 breast cancer cell lines. TRAP1 levels were significantly lower in the MDA468 and MDA231 cell lines, both of which were hormone unresponsive. In hypoxia, TRAP1 was observed to translocate into the nucleus where it associated with Rb. TRAP1 silencing attenuated Rb-E2F1 interaction, resulting in a larger fraction of cells entering into the S phase. Conversely, when TRAP1 was re-expressed in TRAP1-deficient MDA231 breast cancer cells, Rb-E2F1 interaction was enhanced and a smaller S-phase fraction was observed. TRAP1 also appears important in cellular invasion. The invasiveness of MDA231 cells appeared to be attenuated by the restoration of TRAP1, and was possibly due to a reduction in MMP9 levels. Conclusions: TRAP1 appears to modulate tumour behaviour in response to hypoxic stress. Loss of TRAP1 expression increased the proliferative and invasive potential of breast cancer cells, and was associated with a poorer disease-free survival in N2 disease particularly within the first 5 years.
PO40 KI-67 EXPRESSION PREDICTS ADVANCED BREAST CANCER
CLINICAL
OUTCOME
IN
LOCALLY
Naomi Kobayashi, Masahiro Hikichi, Shinsuke Miyajima, Kaori Ushimado, Toshiaki Utsumi Fujita Health University, Dept. of Breast Surgery, Toyoake, Aichi, Japan Goals: The management of locally advanced breast cancer (LABC) requires a combined treatment approach involving surgery, radiation therapy and systemic therapy. Recently, the Ki-67 expression has been considered as an important factor for selecting the additional chemotherapy to endocrine therapy in hormone receptor positive breast cancer. The aim of this study was to examine the prognostic significance of the Ki-67 in LABC. Materials and Methods: One thousand six hundred and twenty patients were diagnosed with breast cancer at our institution from 1992 to 2010. We conducted a retrospective analysis of the 140 patients with LABC (Stage IIIA, IIIB, and IIIC) in this period. The median age of the patients was 56 years with a range of 22 to 92 years old. The expression of Ki-67 was assessed by immunohistochemistry with a monoclonal MIB1 antibody. Ki67 labeling index (LI) was categorized as low (<15%) and high (> or ¼15%) in LABC. Disease-free survival (DFS) curves and over-all survival (OS) curves were estimated using the Kaplan-Meier method and survival comparisons were made with the log-rank test. A multivariate Cox proportional hazards regression model performed in a stepwise fashion was used to determine the prognostic value of each significant variable. The level of significance was taken to be 0.05. IBM SPSS Statistics 19 software package was used for statistical analysis.