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P107 Vasohibin as a potential negative feedback regulator and a newly identified biomarker of angiogenesis in human breast carcinoma
S18
Poster Session I. Biology/Pathology/Basic Research
activity under hypoxic condition than normoxic condition. Furthermore, the small GTPase Ras is also activated in response to hypoxia, which then aids stabilization of HIF-1a, and in turn up-regulates MMP-9 expression. We also demonstrate that MMP-9 is up-regulated concurrently with HIF-1a in tumor tissues from breast cancer patients. Conclusion: These results suggest that HIF-1a promotes cell invasion through a MMP-9-dependent mechanism and that future anti-tumor agents could be used to target HIF-1a and MMP-9. Disclosure of Interest: None Declared
P105
FGFR2 and TP53 work together in breast cancer
E.V. Denisov1 , N.N. Babyshkina1 , M.I. Voevoda2 , N.V. Cherdyntseva1 . 1 Cancer Research Institute, Tomsk, 2 Institute of Internal Medicine, Novosibirsk, Russian Federation Goals: Several studies have identified the association of FGFR2 gene, namely SNPs, with breast cancer (BC) risk, however the mechanism of contribution of FGFR2 polymorphic variants to BC remains unclear up till now. The goal of this study was to compare the distribution of genotype combinations for most significant IVS2+7033A>G (rs1219648) and IVS3+41indel16bp (rs17878362), Ex4+119G>C (rs1042522), IVS6+62G>A (rs1625895) polymorphic variants in FGFR2 and TP53 genes, respectively, between Caucasian patients with sporadic BC (n = 343) and control group (n = 218). Methods: DNA was analyzed for the polymorphism of FGFR2 and TP53 genes by PCR-RFLP. Statistic analysis was performed by STATISTICA 8.0 and Fisher’s exact test. Results: When we applied genotype and statistic analysis, the strong association with BC was attached to only IVS2+7033A>G and Ex4+119G>C genotype combinations. Particularly, the frequency of FGFR2 AA and TP53 GG genotype combination was decreased in BC patients in comparison with control subjects (11.6% vs. 19.7%; p = 0.008), that indicate its protective effect from BC. On the contrary, FGFR2 GG and TP53 GG genotype combination was more often detected in individuals with BC (p = 0.004), pointing to its contribution to BC. When BC groups were stratified depending on woman menopause status, the low frequency of FGFR2 AA and TP53 GG combination was found only in premenopausal women (p = 0.012), whereas the significant increase in combination of GG and GG genotypes was identified in postmenopausal affected subjects only (p = 0.018). Furthermore, we observed the increase in the frequency of FGFR2 GG and TP53 GG genotype combination in patients with multifocal tumors versus subjects who had unifocal tumors (25.9% vs. 11.1%; p = 0.037), that indicates their joint involvement in cancer progression. Conclusion: Taken together, in spite of the fact that the independent association of FGFR2 IVS2+7033A>G and TP53 Ex4+119G>C with BC was shown previously, here, we found that FGFR2 and TP53 variations may make the joint age-related contribution to the disease development. Further researches are required to clarify the mechanisms of their cooperation, however we have some hypothesizes by now. Disclosure of Interest: None Declared
P106
Breast cancer tissue tryptase expression as a new possible antiangiogenic target
M. Ammendola1 , A. Misino2 , R. Patruno3 , V. Goffredo3 , V. Di Lecce4 , T. Martino4 , P. Valerio4 , C.D. Gadaleta3 , G. Ranieri3 . 1 Surgery Unit, Magna Graecia University, Catanzaro, 2 Oncology Unit, National Cancer Institute Giovanni Paolo II, 3 Interventional Radiology Unit with Integrated Section of Medical Oncology, 4 Surgery Unit, Di Venere Hospital, Bari, Italy Goals: Tryptase, a serine protease stored and released from mast cells granules has been identified as a new non-classical angiogenetic factor. Mast cells can release tryptase following c-Kit receptor activation. We have evaluated the correlations among the number of MCs positive to tryptase (MCDPT), the number of c-Kit receptor expressing cells (C-KREC) and microvascular density (MVD) in a series of 88 primary T1−3, N0−2 M0 female breast cancer by means of immunohistochemistry and image analysis methods. Methods: Six-micrometers thick serial sections of formalin-fixed and paraffin-embedded bioptic tumor samples were microwaved at 500 W for 10 min. and treated with a 3% hydrogen peroxide solution. Sections were
Thursday, 17 March 2011 incubated with primary antibodies: anti-tryptase (AA1; Dako, Glostrup, Denmark), anti-c-Kit receptor (A4502; Dako, Glostrup, Denmark) and antiCD34 (QB-END 10; Bio-Optica Milan, Italy). In serial sections MVD, MCDPT and C-KREC were counted by means of image analysis at ×400. Results: Data demonstrated a significantly (r = ranging from 0.71 to 0.91; p: ranging from 0.001 to 0.003 by Pearson’s analysis respectively) correlation between MVD, MCDPT and C-KREC to each other. Conclusion: Published in vitro data suggest that tryptase induce angiogenesis in vascular endothelial cells and breast cancer cells lines. According to these data we shown that MVD, MCDPT and C-KREC paralleled to each other suggesting a role in in vivo breast cancer angiogenesis. In this context several tryptase inhibitors such as gabexate mesilate or nafamostat mesilate might be evaluated in clinical trials as a new antiangiogenetic approach. Disclosure of Interest: None Declared
P107
Vasohibin as a potential negative feedback regulator and a newly identified biomarker of angiogenesis in human breast carcinoma
K. Tamaki1 , H. Sasano2 , M. Miyashita1 , T. Ishida1 , N. Ohuchi1 , N. Tamaki3 . 1 Surgical oncology, 2 Pathology, Tohoku University, Sendai, Miyagi, 3 Breast Surgery, Nahanishi Clinic, Okinawa, Japan Goals: Vasohibin is an identified negative feedback regulator of angiogenesis induced by vascular endothelial growth factor (VEGF)-A and basic fibroblastic growth factor (bFGF). The status of Vasohibin in human malignancies has not been examined in detail. Methods: We examined 251 specimens including 148 cases of invasive ductal carcinoma (IDC), 62 of ductal carcinoma in situ (DCIS), 16 of fibroadenoma, 6 of inflammations, 9 of fibrocystic change and 7 of nonpathological breast tissues. We compared Vasohibin immunoreactivity to that of VEGF-A, bFGF, VEGF receptor 2 (Flk-1) and Ki-67. The correlation of Vasohibin expression with overall survival (OS) and disease free survival (DFS) was also evaluated. We evaluated Ki-67 and CD31 or Vasohibin double-immunostaining for further characterization of neovascularization. We evaluated mRNA expression of Vasohibin and CD31 using a realtime quantitative RT-PCR (QRT-PCR). We also examined the correlation between the expressions of Vasohibin or CD31 and nuclear or histological grades. Results: Vasohibin immunodensity was significantly higher in IDC and inflammations than the other types (P < 0.001). A significant positive correlation was detected between Vasohibin and VEGF-A, bFGF, Flk-1 or Ki-67 (P < 0.001, respectively). There was positive associations between Vasohibin expression and OS (P = 0.004) or DFS (P < 0.001). Results of double-immunostaining demonstrated the ratio of Ki-67 positive cells among Vasohibin (46.5%) was significantly higher than those among CD31 (23.5%). There was no statistically significance in CD31 mRNA expression between DCIS and IDC (P = 0.250), whereas there was a statistically significance in Vasohibin mRNA expression (P = 0.022). Vasohibin expression was correlated with Ki-67, nuclear grade or Van Nuys classification (P < 0.001, respectively). Conclusion: This study indicates that Vasohibin is associated with neovascularization and may play important roles in the regulation of intratumoral angiogenesis. The possible correlation between aggressive biological features in DCIS including increased tumor cell proliferation and the status of neovascularization determined by Vasohibin expression. Disclosure of Interest: None Declared
P108
Isolation of a 320 kDa protein from the eggs of Onchidium (sea slug) that exhibits anticancer activity against breast cancer cell line (T47D)
S. Khodabandeh1 , D. Maleki2 , M. Abdolzadeh3 . 1 Marine Biology, Tarbiat Modares University, Tehran, 2 Oncology Section, Imam Khomeini Hospital, Urmia, 3 Surgery, 15 Khordad Hospital, Tehran, Iran, Islamic Republic of Goals: Pharmacologically active metabolites from marine organisms have extensive use in the treatment of many diseases and serve as compounds of interest both in their natural form and as templates for synthetic modification. Cancer is one of the most lethal diseases in human now, and investigations for finding new anticancer compounds are imperative. Marine invertebrates are interested to identification and isolation of some
Poster Session I. Biology/Pathology/Basic Research
activity under hypoxic condition than normoxic condition. Furthermore, the small GTPase Ras is also activated in response to hypoxia, which then aids stabilization of HIF-1a, and in turn up-regulates MMP-9 expression. We also demonstrate that MMP-9 is up-regulated concurrently with HIF-1a in tumor tissues from breast cancer patients. Conclusion: These results suggest that HIF-1a promotes cell invasion through a MMP-9-dependent mechanism and that future anti-tumor agents could be used to target HIF-1a and MMP-9. Disclosure of Interest: None Declared
P105
FGFR2 and TP53 work together in breast cancer
E.V. Denisov1 , N.N. Babyshkina1 , M.I. Voevoda2 , N.V. Cherdyntseva1 . 1 Cancer Research Institute, Tomsk, 2 Institute of Internal Medicine, Novosibirsk, Russian Federation Goals: Several studies have identified the association of FGFR2 gene, namely SNPs, with breast cancer (BC) risk, however the mechanism of contribution of FGFR2 polymorphic variants to BC remains unclear up till now. The goal of this study was to compare the distribution of genotype combinations for most significant IVS2+7033A>G (rs1219648) and IVS3+41indel16bp (rs17878362), Ex4+119G>C (rs1042522), IVS6+62G>A (rs1625895) polymorphic variants in FGFR2 and TP53 genes, respectively, between Caucasian patients with sporadic BC (n = 343) and control group (n = 218). Methods: DNA was analyzed for the polymorphism of FGFR2 and TP53 genes by PCR-RFLP. Statistic analysis was performed by STATISTICA 8.0 and Fisher’s exact test. Results: When we applied genotype and statistic analysis, the strong association with BC was attached to only IVS2+7033A>G and Ex4+119G>C genotype combinations. Particularly, the frequency of FGFR2 AA and TP53 GG genotype combination was decreased in BC patients in comparison with control subjects (11.6% vs. 19.7%; p = 0.008), that indicate its protective effect from BC. On the contrary, FGFR2 GG and TP53 GG genotype combination was more often detected in individuals with BC (p = 0.004), pointing to its contribution to BC. When BC groups were stratified depending on woman menopause status, the low frequency of FGFR2 AA and TP53 GG combination was found only in premenopausal women (p = 0.012), whereas the significant increase in combination of GG and GG genotypes was identified in postmenopausal affected subjects only (p = 0.018). Furthermore, we observed the increase in the frequency of FGFR2 GG and TP53 GG genotype combination in patients with multifocal tumors versus subjects who had unifocal tumors (25.9% vs. 11.1%; p = 0.037), that indicates their joint involvement in cancer progression. Conclusion: Taken together, in spite of the fact that the independent association of FGFR2 IVS2+7033A>G and TP53 Ex4+119G>C with BC was shown previously, here, we found that FGFR2 and TP53 variations may make the joint age-related contribution to the disease development. Further researches are required to clarify the mechanisms of their cooperation, however we have some hypothesizes by now. Disclosure of Interest: None Declared
P106
Breast cancer tissue tryptase expression as a new possible antiangiogenic target
M. Ammendola1 , A. Misino2 , R. Patruno3 , V. Goffredo3 , V. Di Lecce4 , T. Martino4 , P. Valerio4 , C.D. Gadaleta3 , G. Ranieri3 . 1 Surgery Unit, Magna Graecia University, Catanzaro, 2 Oncology Unit, National Cancer Institute Giovanni Paolo II, 3 Interventional Radiology Unit with Integrated Section of Medical Oncology, 4 Surgery Unit, Di Venere Hospital, Bari, Italy Goals: Tryptase, a serine protease stored and released from mast cells granules has been identified as a new non-classical angiogenetic factor. Mast cells can release tryptase following c-Kit receptor activation. We have evaluated the correlations among the number of MCs positive to tryptase (MCDPT), the number of c-Kit receptor expressing cells (C-KREC) and microvascular density (MVD) in a series of 88 primary T1−3, N0−2 M0 female breast cancer by means of immunohistochemistry and image analysis methods. Methods: Six-micrometers thick serial sections of formalin-fixed and paraffin-embedded bioptic tumor samples were microwaved at 500 W for 10 min. and treated with a 3% hydrogen peroxide solution. Sections were
Thursday, 17 March 2011 incubated with primary antibodies: anti-tryptase (AA1; Dako, Glostrup, Denmark), anti-c-Kit receptor (A4502; Dako, Glostrup, Denmark) and antiCD34 (QB-END 10; Bio-Optica Milan, Italy). In serial sections MVD, MCDPT and C-KREC were counted by means of image analysis at ×400. Results: Data demonstrated a significantly (r = ranging from 0.71 to 0.91; p: ranging from 0.001 to 0.003 by Pearson’s analysis respectively) correlation between MVD, MCDPT and C-KREC to each other. Conclusion: Published in vitro data suggest that tryptase induce angiogenesis in vascular endothelial cells and breast cancer cells lines. According to these data we shown that MVD, MCDPT and C-KREC paralleled to each other suggesting a role in in vivo breast cancer angiogenesis. In this context several tryptase inhibitors such as gabexate mesilate or nafamostat mesilate might be evaluated in clinical trials as a new antiangiogenetic approach. Disclosure of Interest: None Declared
P107
Vasohibin as a potential negative feedback regulator and a newly identified biomarker of angiogenesis in human breast carcinoma
K. Tamaki1 , H. Sasano2 , M. Miyashita1 , T. Ishida1 , N. Ohuchi1 , N. Tamaki3 . 1 Surgical oncology, 2 Pathology, Tohoku University, Sendai, Miyagi, 3 Breast Surgery, Nahanishi Clinic, Okinawa, Japan Goals: Vasohibin is an identified negative feedback regulator of angiogenesis induced by vascular endothelial growth factor (VEGF)-A and basic fibroblastic growth factor (bFGF). The status of Vasohibin in human malignancies has not been examined in detail. Methods: We examined 251 specimens including 148 cases of invasive ductal carcinoma (IDC), 62 of ductal carcinoma in situ (DCIS), 16 of fibroadenoma, 6 of inflammations, 9 of fibrocystic change and 7 of nonpathological breast tissues. We compared Vasohibin immunoreactivity to that of VEGF-A, bFGF, VEGF receptor 2 (Flk-1) and Ki-67. The correlation of Vasohibin expression with overall survival (OS) and disease free survival (DFS) was also evaluated. We evaluated Ki-67 and CD31 or Vasohibin double-immunostaining for further characterization of neovascularization. We evaluated mRNA expression of Vasohibin and CD31 using a realtime quantitative RT-PCR (QRT-PCR). We also examined the correlation between the expressions of Vasohibin or CD31 and nuclear or histological grades. Results: Vasohibin immunodensity was significantly higher in IDC and inflammations than the other types (P < 0.001). A significant positive correlation was detected between Vasohibin and VEGF-A, bFGF, Flk-1 or Ki-67 (P < 0.001, respectively). There was positive associations between Vasohibin expression and OS (P = 0.004) or DFS (P < 0.001). Results of double-immunostaining demonstrated the ratio of Ki-67 positive cells among Vasohibin (46.5%) was significantly higher than those among CD31 (23.5%). There was no statistically significance in CD31 mRNA expression between DCIS and IDC (P = 0.250), whereas there was a statistically significance in Vasohibin mRNA expression (P = 0.022). Vasohibin expression was correlated with Ki-67, nuclear grade or Van Nuys classification (P < 0.001, respectively). Conclusion: This study indicates that Vasohibin is associated with neovascularization and may play important roles in the regulation of intratumoral angiogenesis. The possible correlation between aggressive biological features in DCIS including increased tumor cell proliferation and the status of neovascularization determined by Vasohibin expression. Disclosure of Interest: None Declared
P108
Isolation of a 320 kDa protein from the eggs of Onchidium (sea slug) that exhibits anticancer activity against breast cancer cell line (T47D)
S. Khodabandeh1 , D. Maleki2 , M. Abdolzadeh3 . 1 Marine Biology, Tarbiat Modares University, Tehran, 2 Oncology Section, Imam Khomeini Hospital, Urmia, 3 Surgery, 15 Khordad Hospital, Tehran, Iran, Islamic Republic of Goals: Pharmacologically active metabolites from marine organisms have extensive use in the treatment of many diseases and serve as compounds of interest both in their natural form and as templates for synthetic modification. Cancer is one of the most lethal diseases in human now, and investigations for finding new anticancer compounds are imperative. Marine invertebrates are interested to identification and isolation of some