patrizia casalini | Fond. IRCCS Istituto Nazionale dei Tumori (original) (raw)
Papers by patrizia casalini
Cancer research, Jan 15, 2002
SEL1L, the human orthologue of the Caenorhabditis elegans sel-1 gene, is differentially expressed... more SEL1L, the human orthologue of the Caenorhabditis elegans sel-1 gene, is differentially expressed in breast primary tumors and in normal breast tissues. Analysis of a series of human primary breast carcinomas, using a monoclonal antibody raised against a SEL1L recombinant protein, revealed down-modulation or absence of SEL1L expression in about two-thirds of the tumors as compared with normal breast epithelial cells. Overall survival analysis of breast carcinoma patients indicated a statistically significant correlation between SEL1L down-modulation and poor prognosis. MCF-7, human breast carcinoma cells, were transfected with a construct containing the entire SEL1L cDNA driven by an inducible promoter and showed a dramatic reduction in anchorage-dependent growth and colony formation in soft agar. Growth of the transfected cells in Matrigel, an extracellular matrix rich with laminin, restored colony-formation ability. These results point to the role for SEL1L in breast tumor growth ...
SEL1L, the human orthologue of the Caenorhabditis elegans sel-1 gene, is differentially expressed... more SEL1L, the human orthologue of the Caenorhabditis elegans sel-1 gene, is differentially expressed in breast primary tumors and in normal breast tissues. Analysis of a series of human primary breast carcinomas, using a monoclonal antibody raised against a SEL1L recombinant protein, re- vealed down-modulation or absence of SEL1L expression in about two- thirds of the tumors as compared with normal
Journal of Cellular Physiology, 2015
The tumor-suppressor protein Fhit exerts its functions in the cytoplasm, although some reports su... more The tumor-suppressor protein Fhit exerts its functions in the cytoplasm, although some reports suggest that it may also act in the nucleus. We previously showed that cytosolic Fhit protein levels in cancer cell lines stimulated to proliferate were reduced by proteasomal degradation. Here, we demonstrate that Fhit is physiologically present in the nucleus of breast cancer cell lines and tissues at a low level and that proliferative stimulation increases nuclear levels. Breast cancer cells expressing the FhitY114F mutant, which do not undergo proteasomal degradation, contained mutated Fhit in the nucleus, while cells treated with a proteasome inhibitor accumulated nuclear Fhit during proliferation. Thus, Fhit nuclear shuttling and proteasome degradation phenomena occur independently. When Fhit was coupled to a nuclear localization sequence, the proliferation rate of the transfected cells increased together with levels of proliferation pathway mediators cyclin D1, phospho-MAPK, and phospho-STAT3. Fhit nuclear translocation upon mitogenic stimulation may represent a new regulatory mechanism that allows rapid restoration of Fhit cytoplasmic levels and promotes the proliferation cascade activated by mitogenic stimulation. This article is protected by copyright. All rights reserved.
Molecular Oncology, 2015
Cancer cells within a tumor are functionally heterogeneous and specific subpopulations, defined a... more Cancer cells within a tumor are functionally heterogeneous and specific subpopulations, defined as cancer initiating cells (CICs), are endowed with higher tumor forming potential. The CIC state, however, is not hierarchically stable and conversion of non-CICs to CICs under microenvironment signals might represent a determinant of tumor aggressiveness. How plasticity is regulated at the cellular level is however poorly understood. To identify determinants of plasticity in lung cancer we exposed eight different cell lines to TGFβ1 to induce EMT and stimulate modulation of CD133(+) CICs. We show that response to TGFβ1 treatment is heterogeneous with some cells readily switching to stem cell state (1.5-2 fold CICs increase) and others being unresponsive to stimulation. This response is unrelated to original CICs content or extent of EMT engagement but is tightly dependent on balance between epithelial and mesenchymal features as measured by the ratio of expression of CDH1 (E-cadherin) to SNAI2. Epigenetic modulation of this balance can restore sensitivity of unresponsive models to microenvironmental stimuli, including those elicited by cancer-associated fibroblasts both in vitro and in vivo. In particular, tumors with increased prevalence of cells with features of partial EMT (hybrid epithelial/mesenchymal phenotype) are endowed with the highest plasticity and specific patterns of expression of SNAI2 and CDH1 markers identify a subset of tumors with worse prognosis. In conclusion, here we describe a connection between a hybrid epithelial/mesenchymal phenotype and conversion to stem-cell state in response to external stimuli. These findings have implications for current endeavors to identify tumors with increased plasticity.
Annals of Oncology, 2008
Background: The standardization of the HER2 score and recent changes in therapeutic modalities po... more Background: The standardization of the HER2 score and recent changes in therapeutic modalities points to the need for a reevaluation of the role of HER2 in recently diagnosed breast carcinoma.
Breast Diseases: A Year Book Quarterly, 2008
Results: Onset of LR was significantly linked only to patient's age, with a hazard ratio... more Results: Onset of LR was significantly linked only to patient's age, with a hazard ratio (HR) to relapse reduced by 60% in patients >50 years of age (P = 0.002). Univariate and multivariate Cox analyses in women ≤50 years of age indicated that only expression of hyaluronan in ...
The American Journal of Pathology, 2011
Fibrosis results from inflammatory tissue damage and impaired regeneration. In the context of ble... more Fibrosis results from inflammatory tissue damage and impaired regeneration. In the context of bleomycininduced pulmonary fibrosis, we demonstrated that the matricellular protein termed secreted protein acidic and rich in cysteine (SPARC) distinctly regulates inflammation and collagen deposition, depending on its cellular origin. Reciprocal Sparc ؊/؊ and wild-type (WT) bone marrow chimeras revealed that SPARC expression in host fibroblasts is required and sufficient to induce collagen fibrosis in a proper inflammatory environment. Accordingly, Sparc ؊/؊ >WT chimeras showed exacerbated inflammation and fibrosis due to the inability of Sparc ؊/؊ macrophages to down-regulate tumor necrosis factor production because of impaired responses to tumor growth factor-. Hence, the use of bone marrow cells expressing a dominant-negative form of tumor growth factor- receptor type II under the monocytespecific CD68 promoter, as a decoy, phenocopied Sparc ؊/؊ donor chimeras. Our results point to an unexpected dual role of SPARC in oppositely influencing the outcome of fibrosis.
Journal of The National Cancer Institute, 2003
(1) report findings from the large-scale Italian Randomized Trial of Tamoxifen. Their results sho... more (1) report findings from the large-scale Italian Randomized Trial of Tamoxifen. Their results show that the risk-reducing effects of tamoxifen on high-risk women with breast cancer are most marked for estrogen receptor (ER)-positive tumors. Women at high risk were defined as being ER-positive tumors on the basis of a number of reproductive and hormonal criteria: height of greater than 160 cm, no oophorectomy, menarche by age 13 years, no fullterm pregnancy before age 24 years. This group comprised 702 women (13.0%) of the total cohort (n ס 5395). The results demonstrate that the criteria used to identify the high-risk cohort are valuable, especially in identifying women who may develop ER-positive tumors. These women are particularly suitable for enrollment into chemoprevention studies that interrupt estrogen pathways.
Ejc Supplements, 2010
To evaluate the outcomes of the PET detected hypermetabolic lesions in the lungs of breast cancer... more To evaluate the outcomes of the PET detected hypermetabolic lesions in the lungs of breast cancer patients, as correlated with follow up CT findings and to analyze the PET and CT findings of hypermetabolic lesions in PET-CT for differentiation benign from malignancy.
Breast Cancer Research, 2001
Background: Disruption of the balance between apoptosis and proliferation is considered to be an ... more Background: Disruption of the balance between apoptosis and proliferation is considered to be an important factor in the development and progression of tumor. In this study we determined the in vivo cell kinetics along the spectrum of apparently normal epithelium, hyperplasia, preinvasive lesions and invasive carcinoma, in breast tissues affected by fibrocystic changes in which preinvasive and/or invasive lesions developed, as a model of breast carcinogenesis. Materials and method: A total of 32 areas of apparently normal epithelium and 135 ductal proliferative and neoplastic lesions were studied. More than one epithelial lesion per case was analyzed. The apoptotic index (AI) and the proliferative index (PI) were expressed as the percentage of TUNEL (TdT-mediated dUTP-nick end-labelling) and Ki-67 positive cells, respectively. The proliferative/apoptotic index (P/A) was calculated for each case. Results: Statistical analysis demonstrated significant differences among the tissue groups for both indices (P < 0.0001). The Als and PIs were significantly higher in hyperplasia than in apparently normal epithelium (P = 0.04 and P = 0.0005, respectively), in atypical hyperplasia than in hyperplasia (P = 0.01 and P = 0.04, respectively) and in invasive carcinoma than in in situ carcinoma (P = 0.0001 and P < 0.0001, respectively). The two indices were similar in atypical hyperplasia and in in situ carcinoma. The P/A index increased significantly from normal epithelium to hyperplasia (P = 0.01) and from preinvasive lesions to invasive carcinoma (P = 0.04), whereas it was decreased (NS) from hyperplasia to preinvasive lesions. A strong positive correlation between the Als and the Pls was found (r = 0.83; P < 0.0001). Conclusion: These findings suggest accelerating cell turnover along the continuum of breast carcinogenesis. Atypical hyperplasias and in situ carcinomas might be kinetically similar lesions. In the transition from normal epithelium to hyperplasia and from preinvasive lesions to invasive carcinoma, the net growth of epithelial cells results from a growth imbalance in favour of proliferation. In the transition from hyperplasia to preinvasive lesions there is an imbalance in favour of apoptosis.
Ejc Supplements, 2010
121 Eph/ephrins are hypothesized to be possible mediators of tumour-associated inflammation. The ... more 121 Eph/ephrins are hypothesized to be possible mediators of tumour-associated inflammation. The aim of our study was to analyze the distribution of ephrinB2 and its receptors EphB4 and EphB6 in inflammatory and melanoma cells and to clarify proinflammatory effects due to Eph/ephrin-mediated cell-cell contact. Material and Methods: HL-60 promyelocytes and THP-1 monocytes, differentiated into granulocytes and macrophages, were used as a model for TAIC. Undifferentiated and differentiated cells were co-cultivated with Mel-Juso and A2058 melanoma cells. EphrinB2, EphB4 and EphB6 mRNA expression and protein synthesis was investigated using qRT-PCR and flow cytometry. Secretion of the proinflammatory cytokines IL-6 and TNF-a was analyzed using ELISA. Results: No alteration in gene expression of ephrinB2, EphB4 and EphB6 could be observed during differentiation of HL-60 and THP-1 cells. In contrast, protein synthesis of ephrinB2, EphB4 and EphB6 was two-to threefold higher in HL-60 granulocytes compared to HL-60 promyelocytes and HL-60 macrophages. THP-1 macrophages showed a slightly increased protein synthesis of EphB4 and EphB6 compared to THP-1 monocytes whereas ephrinB2 protein content remained constant. Co-culture of both THP-1 monocytes and macrophages with Mel-Juso cells caused a substantial increment in secretion of proinflammatory cytokines. Co-culture of both HL-60 granulocytes and THP-1 monocytes with A2058 cells did not affect cytokine secretion. By contrast, co-culture of HL-60 macrophages with A2058 cells resulted in increased IL-6 secretion whereas co-culture of THP-1 macrophages with A2058 cells resulted in increased IL-6 secretion but decreased TNF-a release. Conclusions: To our knowledge, mRNA expression and protein synthesis of ephrinB2, EphB4 and EphB6 was investigated for the first time in undifferentiated and differentiated HL-60 and THP-1 cells and, moreover, in Mel-Juso and A2058 melanoma cells. Co-culture of TAIC with melanoma cells resulted in proinflammatory effects. To differentiate the role of various Eph receptors and ephrin ligands in mediation of these effects after direct cell-cell contact of TAIC and melanoma cells selective inhibitors for Eph are applied in ongoing studies.
Breast Cancer Research, 2001
Cancer Immunology Immunotherapy, 1993
In the attempt to define a strategy for screening new monoclonal antibodies (mAb) that could be a... more In the attempt to define a strategy for screening new monoclonal antibodies (mAb) that could be appropriate for clinical application in oncology, we evaluated the suitability of three methods: a direct internalization assay (DIA), an indirect internalization assay (IIA) and an indirect cytotoxicity assay (ICA), by applying them to already selected mAb. The latter were directed against three antigenic systems [38-kDa glycoprotein (gp38), epidermal growth factor receptor, and theneu oncogene product], which, according to their tumor selectivity, could be considered suitable for mAb-guided therapy. The dose-dependent and time-dependent binding, as well as the low intraassay variability, demonstrated the reliability of the three tests. However, a certain degree of inter-assay variability was observed in each one, the highest value being that found when IIA was applied. Furthermore, the degree of variability, as well as the predictability, seemed to be more related to the mAb/antigen (Ag) combination used rather than to the test applied. From the overall data we suggest a procedure to be applied for screening purposes. As a first approach applied to the raw material, ICA is only suitable for screening in the case of an already selected toxin whereas IIA may be helpful to eliminate the true negative mAb. After purification of the relevant mAb a repeated analysis using DIA could allow the selection of true internalizing mAb. However, this second screening should be followed by a further analysis of the fate of the Ag−Ab complex after internalization.
Breast Cancer Research and Treatment, 2002
In a study of invasive breast cancer, multiple correspondence analysis (MCA) revealed clustering ... more In a study of invasive breast cancer, multiple correspondence analysis (MCA) revealed clustering of eight pathobiological variables. Two different phenotypes were distinguished by an index calculated on the basis of the variables (histologic grade, necrosis, lymphoid infiltration, number of mitosis and expression of c-erbB-2, p53, progesterone receptor and Bcl-2). Phenotype A lesions share most of the features of normal breast tissue. Phenotype B looks more malignant, has a higher early recurrence rate and is more frequently seen in younger patients. Our aim was to see if ductal breast carcinoma in situ (DCIS) could be divided into the same phenotypes. One hundred and eighty DCIS were investigated. Association between the eight variables was studied in 2 × 2 models. The phenotype index was calculated by summing weights for the variables in the MCA. All variables were associated, except Bcl-2. DCIS was divided in two phenotypes. Thirty-three tumours were Phenotype A and 147 Phenotype B. The mean age at diagnosis was 65.5 and 58.4 years for Phenotypes A and B, respectively (p = 0.0012). No difference regarding local relapse free survival was seen. Two phenotypes were distinguished in DCIS, similar to invasive breast cancer. In an earlier study, 45% of the invasive cancers were classified as Phenotype B. In this study, 82% of DCIS were Phenotype B. This may indicate that invasive breast cancer of Phenotype B is derived from DCIS of Phenotype B. The distribution of DCIS phenotypes with a small proportion of Phenotype A DCIS may be due to that Phenotype A DCIS is less likely to be detected by mammography, or that some invasive breast cancers of Phenotype A progress to invasiveness without passing the in situ phase.
Annals of Oncology, 2008
Background: The standardization of the HER2 score and recent changes in therapeutic modalities po... more Background: The standardization of the HER2 score and recent changes in therapeutic modalities points to the need for a reevaluation of the role of HER2 in recently diagnosed breast carcinoma.
European Journal of Cancer, 1996
The immunogenicity of the idiotypic portions of two antigrowth factor receptor monoclonal antibod... more The immunogenicity of the idiotypic portions of two antigrowth factor receptor monoclonal antibodies (MAbs) was studied. Immunisation of allogeneic but not syngeneic mice with antihuman epidermal growth factor receptor (EGF-R) MAb MINT5 or anti-HER-2/neu MGR6 MAb elicited a detectable titre of circulating antibodies, particularly when the MAb was coupled with the keyhole limpet haemocyanin and administered together with Freund's adjuvant. The anti-Ab1 response to MAb MINT5 was slightly delayed as compared with the response obtained with MAb MGR6 and was mainly directed to the variable regions. In both cases, all anti-Ab1-positive sera specifically competed with the binding of homologous radiolabelled Ab1 to the relevant EGF-R+ or HER-2/neu+ target cells. Fusion of splenocytes from MINT5-immunised animals failed to produce MAb, whereas cell fusion was successful in generating a paratope-related MAb in the case of MGR6. The anti-MGR6 MAb-produced IdM6.4 inhibited the binding of MAb MGR6 on breast carcinoma cells, suggesting that it recognises an idiotope in or near the antigen combining site, and can be considered useful in the identification and purification of the Abl or its derivatives. We analysed whether a possible recognition of murine EGF-R by MAb MINT5 or a mimicry of EGF by the MAb idiotype prevented or delayed the development of an idiotypic cascade in mice. MINTS inhibited human and murine EGF binding to the human EGF-R, whereas the anti-Ab1 response competed with MINT5 but not with murine EGF binding to A431 human epidermoid carcinoma cells. Moreover, MINT5 did not recognise the murine EGF-R. In a phase I clinical study, no detectable levels of human antimouse antibody response were observed in 5 of the 6 treated cancer patients. The ability of MAb MINT5 to block human EGF-R function, together with its low immunogenicity in patients, raise the possibility of its application in carcinoma immunotherapy.
British Journal of Cancer, 2003
Examination of parity, age at menarche and at menopause by HER2 status in a large series of breas... more Examination of parity, age at menarche and at menopause by HER2 status in a large series of breast carcinomas showed a statistically significant increased-frequency of HER2-positive tumours in lower risk subgroups. The findings suggest a difference in the protective role of hormone-related risk factors between HER2-positive and -negative tumours.
Cancer research, Jan 15, 2002
SEL1L, the human orthologue of the Caenorhabditis elegans sel-1 gene, is differentially expressed... more SEL1L, the human orthologue of the Caenorhabditis elegans sel-1 gene, is differentially expressed in breast primary tumors and in normal breast tissues. Analysis of a series of human primary breast carcinomas, using a monoclonal antibody raised against a SEL1L recombinant protein, revealed down-modulation or absence of SEL1L expression in about two-thirds of the tumors as compared with normal breast epithelial cells. Overall survival analysis of breast carcinoma patients indicated a statistically significant correlation between SEL1L down-modulation and poor prognosis. MCF-7, human breast carcinoma cells, were transfected with a construct containing the entire SEL1L cDNA driven by an inducible promoter and showed a dramatic reduction in anchorage-dependent growth and colony formation in soft agar. Growth of the transfected cells in Matrigel, an extracellular matrix rich with laminin, restored colony-formation ability. These results point to the role for SEL1L in breast tumor growth ...
SEL1L, the human orthologue of the Caenorhabditis elegans sel-1 gene, is differentially expressed... more SEL1L, the human orthologue of the Caenorhabditis elegans sel-1 gene, is differentially expressed in breast primary tumors and in normal breast tissues. Analysis of a series of human primary breast carcinomas, using a monoclonal antibody raised against a SEL1L recombinant protein, re- vealed down-modulation or absence of SEL1L expression in about two- thirds of the tumors as compared with normal
Journal of Cellular Physiology, 2015
The tumor-suppressor protein Fhit exerts its functions in the cytoplasm, although some reports su... more The tumor-suppressor protein Fhit exerts its functions in the cytoplasm, although some reports suggest that it may also act in the nucleus. We previously showed that cytosolic Fhit protein levels in cancer cell lines stimulated to proliferate were reduced by proteasomal degradation. Here, we demonstrate that Fhit is physiologically present in the nucleus of breast cancer cell lines and tissues at a low level and that proliferative stimulation increases nuclear levels. Breast cancer cells expressing the FhitY114F mutant, which do not undergo proteasomal degradation, contained mutated Fhit in the nucleus, while cells treated with a proteasome inhibitor accumulated nuclear Fhit during proliferation. Thus, Fhit nuclear shuttling and proteasome degradation phenomena occur independently. When Fhit was coupled to a nuclear localization sequence, the proliferation rate of the transfected cells increased together with levels of proliferation pathway mediators cyclin D1, phospho-MAPK, and phospho-STAT3. Fhit nuclear translocation upon mitogenic stimulation may represent a new regulatory mechanism that allows rapid restoration of Fhit cytoplasmic levels and promotes the proliferation cascade activated by mitogenic stimulation. This article is protected by copyright. All rights reserved.
Molecular Oncology, 2015
Cancer cells within a tumor are functionally heterogeneous and specific subpopulations, defined a... more Cancer cells within a tumor are functionally heterogeneous and specific subpopulations, defined as cancer initiating cells (CICs), are endowed with higher tumor forming potential. The CIC state, however, is not hierarchically stable and conversion of non-CICs to CICs under microenvironment signals might represent a determinant of tumor aggressiveness. How plasticity is regulated at the cellular level is however poorly understood. To identify determinants of plasticity in lung cancer we exposed eight different cell lines to TGFβ1 to induce EMT and stimulate modulation of CD133(+) CICs. We show that response to TGFβ1 treatment is heterogeneous with some cells readily switching to stem cell state (1.5-2 fold CICs increase) and others being unresponsive to stimulation. This response is unrelated to original CICs content or extent of EMT engagement but is tightly dependent on balance between epithelial and mesenchymal features as measured by the ratio of expression of CDH1 (E-cadherin) to SNAI2. Epigenetic modulation of this balance can restore sensitivity of unresponsive models to microenvironmental stimuli, including those elicited by cancer-associated fibroblasts both in vitro and in vivo. In particular, tumors with increased prevalence of cells with features of partial EMT (hybrid epithelial/mesenchymal phenotype) are endowed with the highest plasticity and specific patterns of expression of SNAI2 and CDH1 markers identify a subset of tumors with worse prognosis. In conclusion, here we describe a connection between a hybrid epithelial/mesenchymal phenotype and conversion to stem-cell state in response to external stimuli. These findings have implications for current endeavors to identify tumors with increased plasticity.
Annals of Oncology, 2008
Background: The standardization of the HER2 score and recent changes in therapeutic modalities po... more Background: The standardization of the HER2 score and recent changes in therapeutic modalities points to the need for a reevaluation of the role of HER2 in recently diagnosed breast carcinoma.
Breast Diseases: A Year Book Quarterly, 2008
Results: Onset of LR was significantly linked only to patient's age, with a hazard ratio... more Results: Onset of LR was significantly linked only to patient's age, with a hazard ratio (HR) to relapse reduced by 60% in patients >50 years of age (P = 0.002). Univariate and multivariate Cox analyses in women ≤50 years of age indicated that only expression of hyaluronan in ...
The American Journal of Pathology, 2011
Fibrosis results from inflammatory tissue damage and impaired regeneration. In the context of ble... more Fibrosis results from inflammatory tissue damage and impaired regeneration. In the context of bleomycininduced pulmonary fibrosis, we demonstrated that the matricellular protein termed secreted protein acidic and rich in cysteine (SPARC) distinctly regulates inflammation and collagen deposition, depending on its cellular origin. Reciprocal Sparc ؊/؊ and wild-type (WT) bone marrow chimeras revealed that SPARC expression in host fibroblasts is required and sufficient to induce collagen fibrosis in a proper inflammatory environment. Accordingly, Sparc ؊/؊ >WT chimeras showed exacerbated inflammation and fibrosis due to the inability of Sparc ؊/؊ macrophages to down-regulate tumor necrosis factor production because of impaired responses to tumor growth factor-. Hence, the use of bone marrow cells expressing a dominant-negative form of tumor growth factor- receptor type II under the monocytespecific CD68 promoter, as a decoy, phenocopied Sparc ؊/؊ donor chimeras. Our results point to an unexpected dual role of SPARC in oppositely influencing the outcome of fibrosis.
Journal of The National Cancer Institute, 2003
(1) report findings from the large-scale Italian Randomized Trial of Tamoxifen. Their results sho... more (1) report findings from the large-scale Italian Randomized Trial of Tamoxifen. Their results show that the risk-reducing effects of tamoxifen on high-risk women with breast cancer are most marked for estrogen receptor (ER)-positive tumors. Women at high risk were defined as being ER-positive tumors on the basis of a number of reproductive and hormonal criteria: height of greater than 160 cm, no oophorectomy, menarche by age 13 years, no fullterm pregnancy before age 24 years. This group comprised 702 women (13.0%) of the total cohort (n ס 5395). The results demonstrate that the criteria used to identify the high-risk cohort are valuable, especially in identifying women who may develop ER-positive tumors. These women are particularly suitable for enrollment into chemoprevention studies that interrupt estrogen pathways.
Ejc Supplements, 2010
To evaluate the outcomes of the PET detected hypermetabolic lesions in the lungs of breast cancer... more To evaluate the outcomes of the PET detected hypermetabolic lesions in the lungs of breast cancer patients, as correlated with follow up CT findings and to analyze the PET and CT findings of hypermetabolic lesions in PET-CT for differentiation benign from malignancy.
Breast Cancer Research, 2001
Background: Disruption of the balance between apoptosis and proliferation is considered to be an ... more Background: Disruption of the balance between apoptosis and proliferation is considered to be an important factor in the development and progression of tumor. In this study we determined the in vivo cell kinetics along the spectrum of apparently normal epithelium, hyperplasia, preinvasive lesions and invasive carcinoma, in breast tissues affected by fibrocystic changes in which preinvasive and/or invasive lesions developed, as a model of breast carcinogenesis. Materials and method: A total of 32 areas of apparently normal epithelium and 135 ductal proliferative and neoplastic lesions were studied. More than one epithelial lesion per case was analyzed. The apoptotic index (AI) and the proliferative index (PI) were expressed as the percentage of TUNEL (TdT-mediated dUTP-nick end-labelling) and Ki-67 positive cells, respectively. The proliferative/apoptotic index (P/A) was calculated for each case. Results: Statistical analysis demonstrated significant differences among the tissue groups for both indices (P < 0.0001). The Als and PIs were significantly higher in hyperplasia than in apparently normal epithelium (P = 0.04 and P = 0.0005, respectively), in atypical hyperplasia than in hyperplasia (P = 0.01 and P = 0.04, respectively) and in invasive carcinoma than in in situ carcinoma (P = 0.0001 and P < 0.0001, respectively). The two indices were similar in atypical hyperplasia and in in situ carcinoma. The P/A index increased significantly from normal epithelium to hyperplasia (P = 0.01) and from preinvasive lesions to invasive carcinoma (P = 0.04), whereas it was decreased (NS) from hyperplasia to preinvasive lesions. A strong positive correlation between the Als and the Pls was found (r = 0.83; P < 0.0001). Conclusion: These findings suggest accelerating cell turnover along the continuum of breast carcinogenesis. Atypical hyperplasias and in situ carcinomas might be kinetically similar lesions. In the transition from normal epithelium to hyperplasia and from preinvasive lesions to invasive carcinoma, the net growth of epithelial cells results from a growth imbalance in favour of proliferation. In the transition from hyperplasia to preinvasive lesions there is an imbalance in favour of apoptosis.
Ejc Supplements, 2010
121 Eph/ephrins are hypothesized to be possible mediators of tumour-associated inflammation. The ... more 121 Eph/ephrins are hypothesized to be possible mediators of tumour-associated inflammation. The aim of our study was to analyze the distribution of ephrinB2 and its receptors EphB4 and EphB6 in inflammatory and melanoma cells and to clarify proinflammatory effects due to Eph/ephrin-mediated cell-cell contact. Material and Methods: HL-60 promyelocytes and THP-1 monocytes, differentiated into granulocytes and macrophages, were used as a model for TAIC. Undifferentiated and differentiated cells were co-cultivated with Mel-Juso and A2058 melanoma cells. EphrinB2, EphB4 and EphB6 mRNA expression and protein synthesis was investigated using qRT-PCR and flow cytometry. Secretion of the proinflammatory cytokines IL-6 and TNF-a was analyzed using ELISA. Results: No alteration in gene expression of ephrinB2, EphB4 and EphB6 could be observed during differentiation of HL-60 and THP-1 cells. In contrast, protein synthesis of ephrinB2, EphB4 and EphB6 was two-to threefold higher in HL-60 granulocytes compared to HL-60 promyelocytes and HL-60 macrophages. THP-1 macrophages showed a slightly increased protein synthesis of EphB4 and EphB6 compared to THP-1 monocytes whereas ephrinB2 protein content remained constant. Co-culture of both THP-1 monocytes and macrophages with Mel-Juso cells caused a substantial increment in secretion of proinflammatory cytokines. Co-culture of both HL-60 granulocytes and THP-1 monocytes with A2058 cells did not affect cytokine secretion. By contrast, co-culture of HL-60 macrophages with A2058 cells resulted in increased IL-6 secretion whereas co-culture of THP-1 macrophages with A2058 cells resulted in increased IL-6 secretion but decreased TNF-a release. Conclusions: To our knowledge, mRNA expression and protein synthesis of ephrinB2, EphB4 and EphB6 was investigated for the first time in undifferentiated and differentiated HL-60 and THP-1 cells and, moreover, in Mel-Juso and A2058 melanoma cells. Co-culture of TAIC with melanoma cells resulted in proinflammatory effects. To differentiate the role of various Eph receptors and ephrin ligands in mediation of these effects after direct cell-cell contact of TAIC and melanoma cells selective inhibitors for Eph are applied in ongoing studies.
Breast Cancer Research, 2001
Cancer Immunology Immunotherapy, 1993
In the attempt to define a strategy for screening new monoclonal antibodies (mAb) that could be a... more In the attempt to define a strategy for screening new monoclonal antibodies (mAb) that could be appropriate for clinical application in oncology, we evaluated the suitability of three methods: a direct internalization assay (DIA), an indirect internalization assay (IIA) and an indirect cytotoxicity assay (ICA), by applying them to already selected mAb. The latter were directed against three antigenic systems [38-kDa glycoprotein (gp38), epidermal growth factor receptor, and theneu oncogene product], which, according to their tumor selectivity, could be considered suitable for mAb-guided therapy. The dose-dependent and time-dependent binding, as well as the low intraassay variability, demonstrated the reliability of the three tests. However, a certain degree of inter-assay variability was observed in each one, the highest value being that found when IIA was applied. Furthermore, the degree of variability, as well as the predictability, seemed to be more related to the mAb/antigen (Ag) combination used rather than to the test applied. From the overall data we suggest a procedure to be applied for screening purposes. As a first approach applied to the raw material, ICA is only suitable for screening in the case of an already selected toxin whereas IIA may be helpful to eliminate the true negative mAb. After purification of the relevant mAb a repeated analysis using DIA could allow the selection of true internalizing mAb. However, this second screening should be followed by a further analysis of the fate of the Ag−Ab complex after internalization.
Breast Cancer Research and Treatment, 2002
In a study of invasive breast cancer, multiple correspondence analysis (MCA) revealed clustering ... more In a study of invasive breast cancer, multiple correspondence analysis (MCA) revealed clustering of eight pathobiological variables. Two different phenotypes were distinguished by an index calculated on the basis of the variables (histologic grade, necrosis, lymphoid infiltration, number of mitosis and expression of c-erbB-2, p53, progesterone receptor and Bcl-2). Phenotype A lesions share most of the features of normal breast tissue. Phenotype B looks more malignant, has a higher early recurrence rate and is more frequently seen in younger patients. Our aim was to see if ductal breast carcinoma in situ (DCIS) could be divided into the same phenotypes. One hundred and eighty DCIS were investigated. Association between the eight variables was studied in 2 × 2 models. The phenotype index was calculated by summing weights for the variables in the MCA. All variables were associated, except Bcl-2. DCIS was divided in two phenotypes. Thirty-three tumours were Phenotype A and 147 Phenotype B. The mean age at diagnosis was 65.5 and 58.4 years for Phenotypes A and B, respectively (p = 0.0012). No difference regarding local relapse free survival was seen. Two phenotypes were distinguished in DCIS, similar to invasive breast cancer. In an earlier study, 45% of the invasive cancers were classified as Phenotype B. In this study, 82% of DCIS were Phenotype B. This may indicate that invasive breast cancer of Phenotype B is derived from DCIS of Phenotype B. The distribution of DCIS phenotypes with a small proportion of Phenotype A DCIS may be due to that Phenotype A DCIS is less likely to be detected by mammography, or that some invasive breast cancers of Phenotype A progress to invasiveness without passing the in situ phase.
Annals of Oncology, 2008
Background: The standardization of the HER2 score and recent changes in therapeutic modalities po... more Background: The standardization of the HER2 score and recent changes in therapeutic modalities points to the need for a reevaluation of the role of HER2 in recently diagnosed breast carcinoma.
European Journal of Cancer, 1996
The immunogenicity of the idiotypic portions of two antigrowth factor receptor monoclonal antibod... more The immunogenicity of the idiotypic portions of two antigrowth factor receptor monoclonal antibodies (MAbs) was studied. Immunisation of allogeneic but not syngeneic mice with antihuman epidermal growth factor receptor (EGF-R) MAb MINT5 or anti-HER-2/neu MGR6 MAb elicited a detectable titre of circulating antibodies, particularly when the MAb was coupled with the keyhole limpet haemocyanin and administered together with Freund's adjuvant. The anti-Ab1 response to MAb MINT5 was slightly delayed as compared with the response obtained with MAb MGR6 and was mainly directed to the variable regions. In both cases, all anti-Ab1-positive sera specifically competed with the binding of homologous radiolabelled Ab1 to the relevant EGF-R+ or HER-2/neu+ target cells. Fusion of splenocytes from MINT5-immunised animals failed to produce MAb, whereas cell fusion was successful in generating a paratope-related MAb in the case of MGR6. The anti-MGR6 MAb-produced IdM6.4 inhibited the binding of MAb MGR6 on breast carcinoma cells, suggesting that it recognises an idiotope in or near the antigen combining site, and can be considered useful in the identification and purification of the Abl or its derivatives. We analysed whether a possible recognition of murine EGF-R by MAb MINT5 or a mimicry of EGF by the MAb idiotype prevented or delayed the development of an idiotypic cascade in mice. MINTS inhibited human and murine EGF binding to the human EGF-R, whereas the anti-Ab1 response competed with MINT5 but not with murine EGF binding to A431 human epidermoid carcinoma cells. Moreover, MINT5 did not recognise the murine EGF-R. In a phase I clinical study, no detectable levels of human antimouse antibody response were observed in 5 of the 6 treated cancer patients. The ability of MAb MINT5 to block human EGF-R function, together with its low immunogenicity in patients, raise the possibility of its application in carcinoma immunotherapy.
British Journal of Cancer, 2003
Examination of parity, age at menarche and at menopause by HER2 status in a large series of breas... more Examination of parity, age at menarche and at menopause by HER2 status in a large series of breast carcinomas showed a statistically significant increased-frequency of HER2-positive tumours in lower risk subgroups. The findings suggest a difference in the protective role of hormone-related risk factors between HER2-positive and -negative tumours.