Human epidermal growth factor receptor 2 (HER2/ERBB2) factor is known to be implicated in many ma... more Human epidermal growth factor receptor 2 (HER2/ERBB2) factor is known to be implicated in many malignancies and the potential of it as a prognostic biomarker was reported years ago. Molecular subtypes of HER2/ERBB2 negative and positive with distinct clinical outcomes have been identified in recent years; however, it is still under investigation for bladder cancer. This study evaluates the biological and prognostic significance of RAD21, RAD50 and BARD1 (homologous recombination biomarkers) mRNA levels with ERBB2 low and high expression to explore their impact on bladder cancer patient survival and cancer aggressiveness. The expression of ERBB2, RAD21, RAD50 and BARD1 mRNA levels was assessed in The Cancer Genome Atlas (TCGA) bladder cancer dataset along with four validation cohorts. Outcome analysis was evaluated using disease-free survival (DFS) and overall survival (OS). Univariate and multivariate analysis were used to evaluate the relationship between RAD21, RAD50, BARD1 and ERBB2 expression and clinicopathological variables. A significant increase in mRNA expression levels of RAD21, RAD50 and BARD1 was noticed in ERBB2-low patients compared to ERBB2-high patients. This overexpression of the homologous recombination repair transcripts was associated with poor outcome in ERBB2-low tumors, not in ERBB2-high tumors. Furthermore, the combined expression of high RAD21/RAD50, high RAD21/BARD1 or high RAD50/BARD1 were significantly associated with worse DFS and a better outcome for those with low co-expression in the ERBB2-low cohort. High expression of either RAD21/RAD50 or RAD21/BARD1 in ERBB2-low cohort associated with higher chance of metastasis. In addition, gene expression of BARD1 alone or in combination with RAD50 acted as an independent prognostic factor for worst survival. The data presented in this study reveal a connection between RAD21, RAD50, BARD1 and ERBB2 and patient survival. Importantly, it provided novel findings and potential prognostic markers, particularly in ERBB2-low bladder cancer. Abbreviations NMIBC Non-muscle invasive bladder cancer MIBC Muscle-invasive bladder cancer HER2/ERBB2 Human epidermal growth factor receptor 2 BRCA1 Breast cancer 1 gene TCGA The Cancer Genome Atlas MSK Memorial Sloan Kettering GEO The Gene Expression Omnibus GO Gene Ontology KEGG Kyoto Encyclopedia of Gene and Genomes DAVID Database for Annotation, Visualization and Integrated Discovery tool
World Journal of Biological Chemistry, Jul 27, 2021
This article examines the hype generated around the term “stem cell”, and the capitalization of t... more This article examines the hype generated around the term “stem cell”, and the capitalization of the stem cell craze by the cosmetic industry. It started by introducing product lines containing active ingredients derived from plant stem cells. Then, evolved to using own cells for skin regeneration and hair loss treatment, and allogenic cells for the manufacturing of stem cell-derived products. This article also discusses the missing links for safe and reliable stem cell applications in cosmetics, and why local regulatory bodies, members of the industry and consumers must all work together to stop the illegitimate use of the “stem cell” good name in unsafe or fraudulent commercial practices.
Journal of Cellular and Molecular Medicine, Sep 3, 2022
Human Epidermal Growth Factor Receptor 2 (HER2) overexpression is considered one of the interesti... more Human Epidermal Growth Factor Receptor 2 (HER2) overexpression is considered one of the interesting prognostic biomarkers in bladder cancer. However, the mechanism of bladder cancer development in relation to HER2 status remains to be elucidated. In this study, we investigated HER2‐Ataxia telangiectasia mutated (ATM) kinase interaction and their impact on patient survival and cancer aggressiveness. Using the Cancer Genome Atlas (TCGA) cohorts, we demonstrated that ATM expression (protein/mRNA) is increased in HER2 deficient compared with proficient HER2 patients. This finding was then validated using the Gene Expression Omnibus database (GEO). Correlation analysis (using low expression vs high expression as a discriminator) revealed a significant association of ATM low and HER2 high status with several clinicopathological variables such as high tumour grade, late disease stage and tumour shape. Kaplan–Meier survival analysis indicated that ATM low and HER2 high is a powerful prognosticator of both overall survival (OS) and disease‐free survival (DFS). Furthermore, using bioinformatics and protein/protein interaction analyses, we identified 66 putative overlapping proteins with direct link between HER2 and ATM most of which are functionally involved in transcription regulation, apoptotic process and cell proliferation. Interestingly, the results showed that these proteins are strongly linked with PI3K‐Akt pathway, p53 pathway and microRNAs in cancer. Altogether, our data pinpoint an important biological role of the interconnection between HER2 and ATM. The latter appear to be an independent prognostic biomarker and may serve as targets to develop novel combination therapies to improve the outcome of patients with bladder cancer.
The development and progression of any tumour is not only determined by the corresponding cancer ... more The development and progression of any tumour is not only determined by the corresponding cancer cells but also by the microenvironment of the tumour. This includes an orchestrated network of interacting cell types such as Mesenchymal Stem Cells (MSCs), immune cells, and endothelial cells, through the extracellular matrix and different soluble factors. Breast Cancer (BC) is a leading cause of death in women around the world. BC is not a single disease, but it is a collection of diseases that have different histopathological features, genetic and genomic variability, and the significant prognostic factors varied by diagnosis. MSCs derived from adipose tissue, bone marrow, placenta and other tissue, are multipotent adult cells with potential to treat human diseases such as cancer. A major development has been introduced in defining cellular hierarchy and the niche of stem cell in the human mammary gland. For decades autologous fat grafting has been suggested to be used after mastectomy for reconstructive purposes (restore form and anatomy). Additionally, adipose fat has the inherent advantage of being autologous tissue. It is considered to be the best natural-appearing filler; nevertheless given its unpredictable engraftment and rates of retention, it lacks reliability. Presently, stem cells have become the targets of BC therapy, although the investigations are mostly on a basic stage level. In this review we discuss the double-edged sword of MSC in BC.
FABP7 is a member of the multi-gene fatty acid binding protein family. It is expressed in the mam... more FABP7 is a member of the multi-gene fatty acid binding protein family. It is expressed in the mammary gland and has been shown to function as inhibitor of proliferation of breast tumour cells and to promote differentiation through the JAK/Stat pathway. Cytoplasmic FABP7 expression has been shown to be associated with a favourable prognosis of basal-like breast cancer. In other tissues, varying sub-cellular localization of FABP7 between the nucleus and cytoplasm has been observed. Tissue microarray preparations of well-characterized series of 1,249 unselected and 245 ER-negative invasive breast cancers with a long-term follow-up were investigated in this study to assess the biological and clinical significance of FABP7 sub-cellular localization using immunohistochemistry. Both nuclear and cytoplasmic FABP7 were observed. Nuclear FABP7 was associated with high histologic grade, mitotic frequency, pleomorphism and stage, in addition to basal phenotype (BP) and triple-negative (TN) phenotype. Nuclear FABP7 expression showed an association with expression of markers associated with proliferation and cell-cycle control including Ki67, p53 and p21; however, cytoplasmic FABP7 was associated only with Ki67 and P53 (P = 0.001, < 0.001 respectively). Interestingly, in multivariate analysis, nuclear FABP7 expression in BP was significantly associated with longer DFI (P = 0.025) independent of cytoplasmic expression. Tumours with only nuclear positive FABP7 expression had significantly better prognosis than those with only cytoplasmic expression. This is the first study elucidating the sub-cellular localization of FABP7 in a large series of breast cancer cases. Our observations demonstrate the considerable heterogeneity in expression patterns of FABP7 within breast cancer that relates to differences in biological behaviour especially in basal-like breast cancer. Further investigation of the biology of FABP7 in breast cancer is warranted.
Breast Cancer Research and Treatment, Apr 16, 2013
Breast cancer is recognised to be a heterogeneous disease and the second most common cause of mor... more Breast cancer is recognised to be a heterogeneous disease and the second most common cause of morbidity and mortality worldwide in women. Basal-like breast cancer (BLBC) is associated with aggressive characteristics including development of recurrent disease and reduced survival. BLBC has been defined in some studies as tumours lacking both oestrogen receptor and progesterone receptor protein expression. Gene expression studies have shown that these tumours are also associated with expression of basal-type cytokeratins, the phenotypic patterns of basal cytokeratin expression in BLBC have not been widely studied. A well-characterised series of 995 invasive breast cancers with a long-term follow up were investigated using immunohistochemical staining for four basal cytokeratins (CK5, CK5/6, CK14 and CK17). The data were analysed using univariate and clustering analysis. As a result BLBC, as defined by negativity for ER and HER2 showed variable positivity for basal cytokeratin expression: 61.7 % CK5, 50.5 % CK5/6, 24.2 % CK14 and 23 % CK17. These characteristics were associated with poor outcome characteristics including high histological grade, mitosis, pleomorphism and tumour size [1.5 cm. CK5 positivity was more associated with ER-, PgR-, TN and double ER-PgR-, than the other cytokeratins. Four different clusters of basal cytokeratin expression patterns were identified: (1) negativity for all basal cytokeratins, (2) CK5 ? /CK17-, (3) CK5-/CK17 ? and (4) CK5 ? /CK17 ?. These patterns of basal cytokeratin expression associated with differences in patient outcome, clusters 1 and 3 showed better outcomes than cluster 4 and 2, with cluster 2 having the poorest prognosis. In conclusion, four basal cytokeratin expression patterns were identified in human breast cancer using unsupervised clustering analysis and these patterns are associated with differences in patient outcome.
Supplementary Tables S1-S9: Supplementary Table S1: Clinicopathological characteristics in the ME... more Supplementary Tables S1-S9: Supplementary Table S1: Clinicopathological characteristics in the METABRIC cohort Supplementary Table S2: External validation cohorts (pooled n = 2413). Supplementary Table S3: Clinicopathological characteristics of Nottingham cohort Supplementary Table S4: Antigens, primary antibodies, clone, source, optimal dilution and scoring system used for each immunohistochemical marker Supplementary Table S5: BLM (nuclear protein expression) in breast cancer Supplementary Table S6: BLM (cytoplasmic protein expression) in breast cancer Supplementary Table S7: BLM (nuclear and cytoplasmic protein co-expression) in breast cancer Supplementary Table S8. BLM - Rad51 nuclear co-expression and breast cancer Supplementary Table S9: Multivariate analysis in Nottingham cohort.
Transcriptional activation by estrogen receptor (ER) is a key step to breast oncogenesis. Given p... more Transcriptional activation by estrogen receptor (ER) is a key step to breast oncogenesis. Given previous findings that ADA3 is a critical component of HAT complexes that regulate ER function and evidence that overexpression of other ER coactivators such as SRC-3 is associated with clinical outcomes in breast cancer, the current study was designed to assess the potential significance of ADA3 expression/ localization in human breast cancer patients. In this study, we analyzed ADA3 expression in breast cancer tissue specimens and assessed the correlation of ADA3 staining with cancer progression and patient outcome. Tissue microarrays prepared from large series of breast cancer patients with long-term follow-ups were stained with anti-ADA3 monoclonal antibody using immunohistochemistry. Samples were analyzed for ADA3 expression followed by correlation with various clinicopathological parameters and patients' outcomes. We report that breast cancer specimens show predominant nuclear, cytoplasmic, or mixed nuclear ? cytoplasmic ADA3 staining patterns. Predominant nuclear ADA3 staining correlated with ER? status. While predominant cytoplasmic ADA3 staining negatively correlated with ER? status, but positively correlated with ErbB2, EGFR, and Ki67. Furthermore, a positive correlation of cytoplasmic ADA3 was observed with higher histological grade, mitotic counts, Nottingham Prognostic Index, and positive vascular invasion. Patients with nuclear ADA3 and ER positivity have better breast cancer specific survival and distant metastasis free survival. Significantly, cytoplasmic expression of ADA3 showed a strong positive association with reduced BCSS and DMFS in ErbB2?/EGFR? patients. Although in multivariate analyses ADA3 expression was not an independent marker of survival, predominant nuclear ADA3 staining in breast cancer tissues correlates with ER? expression and together serves as a marker of good prognosis, whereas predominant cytoplasmic ADA3 expression correlates with ErbB2?/EGFR? expression and together is a marker of poor prognosis. Thus, ADA3 cytoplasmic localization together with ErbB2?/ Electronic supplementary material The online version of this article (
Breast cancer is the most common type of cancer and second leading cause of cancer-related deaths... more Breast cancer is the most common type of cancer and second leading cause of cancer-related deaths in women in the United States. Our laboratory has identified the protein hEcd (human ortholog of Drosophila Ecdysoneless) as a novel regulator of cell cycle. Previous studies showed that Ecd regulates cell cycle by regulating the Rb-E2F pathway. Given the dysregulation of the cell cycle machinery in cancer, we examined expression of Ecd in normal, benign, ductal carcinoma in situ (DCIS) and infiltrating ductal carcinoma (IDCs) of breast. In first cohort, we showed while normal and hyperplasia of breast barely showed Ecd expression, high Ecd expression was observed in DCIS and IDC patient tissue specimens. In this cohort of 104 IDC patents, Ecd expression was positively associated with higher grade (p=0.04).We then analyzed Ecd expression in a second larger cohort (954) and observed similar results, where increased Ecd expression was associated with tumors of higher histological grade (p=0.013), mitotic count (p=0.032), and Nottingham Prognostic Index score (p=0.014). Notably, Ecd expression was positively associated with HER2/neu (p=0.002) overexpression. Significantly, a positive association between Ecd expression and shorter breast cancer specific survival (BCSS) (p=0.008) and disease-free survival (DFS) (p=0.003) was observed in HER2/neu overexpressing patients. Taken together, our results demonstrate Ecd expression as a novel marker for breast cancer progression that predicts tumor progression and the clinical outcome in breast cancer patients. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4593. doi:1538-7445.AM2012-4593
Background: Checkpoint kinase1 (CHK1), which is a key component of DNA-damage-activated checkpoin... more Background: Checkpoint kinase1 (CHK1), which is a key component of DNA-damage-activated checkpoint signalling response, may have a role in breast cancer (BC) pathogenesis and influence response to chemotherapy. This study investigated the clinicopathological significance of phosphorylated CHK1 (pCHK1) protein in BC. Method: pCHK1 protein expression was assessed using immunohistochemistry in a large, well-characterized annotated series of early-stage primary operable invasive BC prepared as tissue microarray (n ¼ 1200). Result: pCHK1 showed nuclear and/or cytoplasmic expression. Tumours with nuclear expression showed positive associations with favourable prognostic features such as lower grade, lower mitotic activity, expression of hormone receptor and lack of expression of KI67 and PI3K (Po0.001). On the other hand, cytoplasmic expression was associated with features of poor prognosis such as higher grade, triple-negative phenotype and expression of KI67, p53, AKT and PI3K. pCHK1 expression showed an association with DNA damage response (ATM, RAD51, BRCA1, KU70/KU80, DNA-PKCa and BARD1) and sumoylation (UBC9 and PIASg) biomarkers. Subcellular localisation of pCHK1 was associated with the expression of the nuclear transport protein KPNA2. Positive nuclear expression predicted better survival outcome in patients who did not receive chemotherapy in the whole series and in ER-positive tumours. In ER-negative and triple-negative subgroups, nuclear pCHK1 predicted shorter survival in patients who received cyclophosphamide, methotrexate and 5-florouracil chemotherapy. Conclusions: Our data suggest that pCHK1 may have prognostic and predictive significance in BC. Subcellular localisation of pCHK1 protein is related to its function. Systemic therapy targeted at disrupting DNA damage response (DDR) in solid tumours is attracting attention and it is likely to expand in the coming years. A complex series of checkpoint pathways are recognised to have important roles in the repair of any DNA damage (Niida and Nakanishi, 2006). This mechanism is mediated through an array of DDR proteins that when activated are involved in arrest of cell cycle to allow DNA repair process to proceed, thus preventing DNA replication and mitosis in the presence of unrepaired damage; otherwise apoptosis or mutagenesis will occur (Bartek and Lukas, 2003; Dai and Grant, 2010).
Breast Cancer Research and Treatment, Apr 18, 2014
The definition of Luminal-B subclass of breast cancer (BC) varies in literature. In this study, w... more The definition of Luminal-B subclass of breast cancer (BC) varies in literature. In this study, we have compared the proliferation status; assessed using KI67 labeling index (KI67-LI), and HER2-expression in estrogen receptor positive (ER?) BC to assess their impact on the biological and clinical characteristics of luminal-BC. 1547 (73.8 %) well-characterized clinically annotated stage I-III ER ? BC were assessed for expression of KI67, HER2 (ASCO guidelines), and a large panel of relevant biomarkers (no = 37). 46.3 % of the cases show high KI67-LI ([13 %) and 8.4 % show HER2? and both markers are positively associated with younger age, higher tumor grade and poorer outcome. High KI67-LI and HER2? are associated with upregulation of ER-coactivators and proliferation-related markers and with downregulation of good prognostic markers. High KI67-LI is associated with larger size, advanced stage, and lymphovascular invasion (LVI) and with downregulation of luminal-enriched and DNA-damage repair markers. In contrast, HER2? is associated with upregulation of ERregulated proteins and E-cadherin. When analysis is restricted to high KI67-LI subgroup, HER2? shows an association with upregulation of differentiation-associated proteins and E-cadherin. Conversely, within HER2? class, high KI67-LI maintains its association with downregulation of differentiation-associated/luminal-enriched proteins. Outcome analyses indicate that both markers are independently associated with shorter survival but HER2? is associated with a worse outcome. Although both are associated with high proliferation and poor prognosis within ER ? BC, HER2? is less frequent than high KI67-LI. Unlike KI67, HER2 seems to independently drive the aggressive behavior of ER? tumors without downregulation of luminal proteins.
Human epidermal growth factor receptor 2 (HER2/ERBB2) factor is known to be implicated in many ma... more Human epidermal growth factor receptor 2 (HER2/ERBB2) factor is known to be implicated in many malignancies and the potential of it as a prognostic biomarker was reported years ago. Molecular subtypes of HER2/ERBB2 negative and positive with distinct clinical outcomes have been identified in recent years; however, it is still under investigation for bladder cancer. This study evaluates the biological and prognostic significance of RAD21, RAD50 and BARD1 (homologous recombination biomarkers) mRNA levels with ERBB2 low and high expression to explore their impact on bladder cancer patient survival and cancer aggressiveness. The expression of ERBB2, RAD21, RAD50 and BARD1 mRNA levels was assessed in The Cancer Genome Atlas (TCGA) bladder cancer dataset along with four validation cohorts. Outcome analysis was evaluated using disease-free survival (DFS) and overall survival (OS). Univariate and multivariate analysis were used to evaluate the relationship between RAD21, RAD50, BARD1 and ERBB2 expression and clinicopathological variables. A significant increase in mRNA expression levels of RAD21, RAD50 and BARD1 was noticed in ERBB2-low patients compared to ERBB2-high patients. This overexpression of the homologous recombination repair transcripts was associated with poor outcome in ERBB2-low tumors, not in ERBB2-high tumors. Furthermore, the combined expression of high RAD21/RAD50, high RAD21/BARD1 or high RAD50/BARD1 were significantly associated with worse DFS and a better outcome for those with low co-expression in the ERBB2-low cohort. High expression of either RAD21/RAD50 or RAD21/BARD1 in ERBB2-low cohort associated with higher chance of metastasis. In addition, gene expression of BARD1 alone or in combination with RAD50 acted as an independent prognostic factor for worst survival. The data presented in this study reveal a connection between RAD21, RAD50, BARD1 and ERBB2 and patient survival. Importantly, it provided novel findings and potential prognostic markers, particularly in ERBB2-low bladder cancer. Abbreviations NMIBC Non-muscle invasive bladder cancer MIBC Muscle-invasive bladder cancer HER2/ERBB2 Human epidermal growth factor receptor 2 BRCA1 Breast cancer 1 gene TCGA The Cancer Genome Atlas MSK Memorial Sloan Kettering GEO The Gene Expression Omnibus GO Gene Ontology KEGG Kyoto Encyclopedia of Gene and Genomes DAVID Database for Annotation, Visualization and Integrated Discovery tool
World Journal of Biological Chemistry, Jul 27, 2021
This article examines the hype generated around the term “stem cell”, and the capitalization of t... more This article examines the hype generated around the term “stem cell”, and the capitalization of the stem cell craze by the cosmetic industry. It started by introducing product lines containing active ingredients derived from plant stem cells. Then, evolved to using own cells for skin regeneration and hair loss treatment, and allogenic cells for the manufacturing of stem cell-derived products. This article also discusses the missing links for safe and reliable stem cell applications in cosmetics, and why local regulatory bodies, members of the industry and consumers must all work together to stop the illegitimate use of the “stem cell” good name in unsafe or fraudulent commercial practices.
Journal of Cellular and Molecular Medicine, Sep 3, 2022
Human Epidermal Growth Factor Receptor 2 (HER2) overexpression is considered one of the interesti... more Human Epidermal Growth Factor Receptor 2 (HER2) overexpression is considered one of the interesting prognostic biomarkers in bladder cancer. However, the mechanism of bladder cancer development in relation to HER2 status remains to be elucidated. In this study, we investigated HER2‐Ataxia telangiectasia mutated (ATM) kinase interaction and their impact on patient survival and cancer aggressiveness. Using the Cancer Genome Atlas (TCGA) cohorts, we demonstrated that ATM expression (protein/mRNA) is increased in HER2 deficient compared with proficient HER2 patients. This finding was then validated using the Gene Expression Omnibus database (GEO). Correlation analysis (using low expression vs high expression as a discriminator) revealed a significant association of ATM low and HER2 high status with several clinicopathological variables such as high tumour grade, late disease stage and tumour shape. Kaplan–Meier survival analysis indicated that ATM low and HER2 high is a powerful prognosticator of both overall survival (OS) and disease‐free survival (DFS). Furthermore, using bioinformatics and protein/protein interaction analyses, we identified 66 putative overlapping proteins with direct link between HER2 and ATM most of which are functionally involved in transcription regulation, apoptotic process and cell proliferation. Interestingly, the results showed that these proteins are strongly linked with PI3K‐Akt pathway, p53 pathway and microRNAs in cancer. Altogether, our data pinpoint an important biological role of the interconnection between HER2 and ATM. The latter appear to be an independent prognostic biomarker and may serve as targets to develop novel combination therapies to improve the outcome of patients with bladder cancer.
The development and progression of any tumour is not only determined by the corresponding cancer ... more The development and progression of any tumour is not only determined by the corresponding cancer cells but also by the microenvironment of the tumour. This includes an orchestrated network of interacting cell types such as Mesenchymal Stem Cells (MSCs), immune cells, and endothelial cells, through the extracellular matrix and different soluble factors. Breast Cancer (BC) is a leading cause of death in women around the world. BC is not a single disease, but it is a collection of diseases that have different histopathological features, genetic and genomic variability, and the significant prognostic factors varied by diagnosis. MSCs derived from adipose tissue, bone marrow, placenta and other tissue, are multipotent adult cells with potential to treat human diseases such as cancer. A major development has been introduced in defining cellular hierarchy and the niche of stem cell in the human mammary gland. For decades autologous fat grafting has been suggested to be used after mastectomy for reconstructive purposes (restore form and anatomy). Additionally, adipose fat has the inherent advantage of being autologous tissue. It is considered to be the best natural-appearing filler; nevertheless given its unpredictable engraftment and rates of retention, it lacks reliability. Presently, stem cells have become the targets of BC therapy, although the investigations are mostly on a basic stage level. In this review we discuss the double-edged sword of MSC in BC.
FABP7 is a member of the multi-gene fatty acid binding protein family. It is expressed in the mam... more FABP7 is a member of the multi-gene fatty acid binding protein family. It is expressed in the mammary gland and has been shown to function as inhibitor of proliferation of breast tumour cells and to promote differentiation through the JAK/Stat pathway. Cytoplasmic FABP7 expression has been shown to be associated with a favourable prognosis of basal-like breast cancer. In other tissues, varying sub-cellular localization of FABP7 between the nucleus and cytoplasm has been observed. Tissue microarray preparations of well-characterized series of 1,249 unselected and 245 ER-negative invasive breast cancers with a long-term follow-up were investigated in this study to assess the biological and clinical significance of FABP7 sub-cellular localization using immunohistochemistry. Both nuclear and cytoplasmic FABP7 were observed. Nuclear FABP7 was associated with high histologic grade, mitotic frequency, pleomorphism and stage, in addition to basal phenotype (BP) and triple-negative (TN) phenotype. Nuclear FABP7 expression showed an association with expression of markers associated with proliferation and cell-cycle control including Ki67, p53 and p21; however, cytoplasmic FABP7 was associated only with Ki67 and P53 (P = 0.001, < 0.001 respectively). Interestingly, in multivariate analysis, nuclear FABP7 expression in BP was significantly associated with longer DFI (P = 0.025) independent of cytoplasmic expression. Tumours with only nuclear positive FABP7 expression had significantly better prognosis than those with only cytoplasmic expression. This is the first study elucidating the sub-cellular localization of FABP7 in a large series of breast cancer cases. Our observations demonstrate the considerable heterogeneity in expression patterns of FABP7 within breast cancer that relates to differences in biological behaviour especially in basal-like breast cancer. Further investigation of the biology of FABP7 in breast cancer is warranted.
Breast Cancer Research and Treatment, Apr 16, 2013
Breast cancer is recognised to be a heterogeneous disease and the second most common cause of mor... more Breast cancer is recognised to be a heterogeneous disease and the second most common cause of morbidity and mortality worldwide in women. Basal-like breast cancer (BLBC) is associated with aggressive characteristics including development of recurrent disease and reduced survival. BLBC has been defined in some studies as tumours lacking both oestrogen receptor and progesterone receptor protein expression. Gene expression studies have shown that these tumours are also associated with expression of basal-type cytokeratins, the phenotypic patterns of basal cytokeratin expression in BLBC have not been widely studied. A well-characterised series of 995 invasive breast cancers with a long-term follow up were investigated using immunohistochemical staining for four basal cytokeratins (CK5, CK5/6, CK14 and CK17). The data were analysed using univariate and clustering analysis. As a result BLBC, as defined by negativity for ER and HER2 showed variable positivity for basal cytokeratin expression: 61.7 % CK5, 50.5 % CK5/6, 24.2 % CK14 and 23 % CK17. These characteristics were associated with poor outcome characteristics including high histological grade, mitosis, pleomorphism and tumour size [1.5 cm. CK5 positivity was more associated with ER-, PgR-, TN and double ER-PgR-, than the other cytokeratins. Four different clusters of basal cytokeratin expression patterns were identified: (1) negativity for all basal cytokeratins, (2) CK5 ? /CK17-, (3) CK5-/CK17 ? and (4) CK5 ? /CK17 ?. These patterns of basal cytokeratin expression associated with differences in patient outcome, clusters 1 and 3 showed better outcomes than cluster 4 and 2, with cluster 2 having the poorest prognosis. In conclusion, four basal cytokeratin expression patterns were identified in human breast cancer using unsupervised clustering analysis and these patterns are associated with differences in patient outcome.
Supplementary Tables S1-S9: Supplementary Table S1: Clinicopathological characteristics in the ME... more Supplementary Tables S1-S9: Supplementary Table S1: Clinicopathological characteristics in the METABRIC cohort Supplementary Table S2: External validation cohorts (pooled n = 2413). Supplementary Table S3: Clinicopathological characteristics of Nottingham cohort Supplementary Table S4: Antigens, primary antibodies, clone, source, optimal dilution and scoring system used for each immunohistochemical marker Supplementary Table S5: BLM (nuclear protein expression) in breast cancer Supplementary Table S6: BLM (cytoplasmic protein expression) in breast cancer Supplementary Table S7: BLM (nuclear and cytoplasmic protein co-expression) in breast cancer Supplementary Table S8. BLM - Rad51 nuclear co-expression and breast cancer Supplementary Table S9: Multivariate analysis in Nottingham cohort.
Transcriptional activation by estrogen receptor (ER) is a key step to breast oncogenesis. Given p... more Transcriptional activation by estrogen receptor (ER) is a key step to breast oncogenesis. Given previous findings that ADA3 is a critical component of HAT complexes that regulate ER function and evidence that overexpression of other ER coactivators such as SRC-3 is associated with clinical outcomes in breast cancer, the current study was designed to assess the potential significance of ADA3 expression/ localization in human breast cancer patients. In this study, we analyzed ADA3 expression in breast cancer tissue specimens and assessed the correlation of ADA3 staining with cancer progression and patient outcome. Tissue microarrays prepared from large series of breast cancer patients with long-term follow-ups were stained with anti-ADA3 monoclonal antibody using immunohistochemistry. Samples were analyzed for ADA3 expression followed by correlation with various clinicopathological parameters and patients' outcomes. We report that breast cancer specimens show predominant nuclear, cytoplasmic, or mixed nuclear ? cytoplasmic ADA3 staining patterns. Predominant nuclear ADA3 staining correlated with ER? status. While predominant cytoplasmic ADA3 staining negatively correlated with ER? status, but positively correlated with ErbB2, EGFR, and Ki67. Furthermore, a positive correlation of cytoplasmic ADA3 was observed with higher histological grade, mitotic counts, Nottingham Prognostic Index, and positive vascular invasion. Patients with nuclear ADA3 and ER positivity have better breast cancer specific survival and distant metastasis free survival. Significantly, cytoplasmic expression of ADA3 showed a strong positive association with reduced BCSS and DMFS in ErbB2?/EGFR? patients. Although in multivariate analyses ADA3 expression was not an independent marker of survival, predominant nuclear ADA3 staining in breast cancer tissues correlates with ER? expression and together serves as a marker of good prognosis, whereas predominant cytoplasmic ADA3 expression correlates with ErbB2?/EGFR? expression and together is a marker of poor prognosis. Thus, ADA3 cytoplasmic localization together with ErbB2?/ Electronic supplementary material The online version of this article (
Breast cancer is the most common type of cancer and second leading cause of cancer-related deaths... more Breast cancer is the most common type of cancer and second leading cause of cancer-related deaths in women in the United States. Our laboratory has identified the protein hEcd (human ortholog of Drosophila Ecdysoneless) as a novel regulator of cell cycle. Previous studies showed that Ecd regulates cell cycle by regulating the Rb-E2F pathway. Given the dysregulation of the cell cycle machinery in cancer, we examined expression of Ecd in normal, benign, ductal carcinoma in situ (DCIS) and infiltrating ductal carcinoma (IDCs) of breast. In first cohort, we showed while normal and hyperplasia of breast barely showed Ecd expression, high Ecd expression was observed in DCIS and IDC patient tissue specimens. In this cohort of 104 IDC patents, Ecd expression was positively associated with higher grade (p=0.04).We then analyzed Ecd expression in a second larger cohort (954) and observed similar results, where increased Ecd expression was associated with tumors of higher histological grade (p=0.013), mitotic count (p=0.032), and Nottingham Prognostic Index score (p=0.014). Notably, Ecd expression was positively associated with HER2/neu (p=0.002) overexpression. Significantly, a positive association between Ecd expression and shorter breast cancer specific survival (BCSS) (p=0.008) and disease-free survival (DFS) (p=0.003) was observed in HER2/neu overexpressing patients. Taken together, our results demonstrate Ecd expression as a novel marker for breast cancer progression that predicts tumor progression and the clinical outcome in breast cancer patients. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4593. doi:1538-7445.AM2012-4593
Background: Checkpoint kinase1 (CHK1), which is a key component of DNA-damage-activated checkpoin... more Background: Checkpoint kinase1 (CHK1), which is a key component of DNA-damage-activated checkpoint signalling response, may have a role in breast cancer (BC) pathogenesis and influence response to chemotherapy. This study investigated the clinicopathological significance of phosphorylated CHK1 (pCHK1) protein in BC. Method: pCHK1 protein expression was assessed using immunohistochemistry in a large, well-characterized annotated series of early-stage primary operable invasive BC prepared as tissue microarray (n ¼ 1200). Result: pCHK1 showed nuclear and/or cytoplasmic expression. Tumours with nuclear expression showed positive associations with favourable prognostic features such as lower grade, lower mitotic activity, expression of hormone receptor and lack of expression of KI67 and PI3K (Po0.001). On the other hand, cytoplasmic expression was associated with features of poor prognosis such as higher grade, triple-negative phenotype and expression of KI67, p53, AKT and PI3K. pCHK1 expression showed an association with DNA damage response (ATM, RAD51, BRCA1, KU70/KU80, DNA-PKCa and BARD1) and sumoylation (UBC9 and PIASg) biomarkers. Subcellular localisation of pCHK1 was associated with the expression of the nuclear transport protein KPNA2. Positive nuclear expression predicted better survival outcome in patients who did not receive chemotherapy in the whole series and in ER-positive tumours. In ER-negative and triple-negative subgroups, nuclear pCHK1 predicted shorter survival in patients who received cyclophosphamide, methotrexate and 5-florouracil chemotherapy. Conclusions: Our data suggest that pCHK1 may have prognostic and predictive significance in BC. Subcellular localisation of pCHK1 protein is related to its function. Systemic therapy targeted at disrupting DNA damage response (DDR) in solid tumours is attracting attention and it is likely to expand in the coming years. A complex series of checkpoint pathways are recognised to have important roles in the repair of any DNA damage (Niida and Nakanishi, 2006). This mechanism is mediated through an array of DDR proteins that when activated are involved in arrest of cell cycle to allow DNA repair process to proceed, thus preventing DNA replication and mitosis in the presence of unrepaired damage; otherwise apoptosis or mutagenesis will occur (Bartek and Lukas, 2003; Dai and Grant, 2010).
Breast Cancer Research and Treatment, Apr 18, 2014
The definition of Luminal-B subclass of breast cancer (BC) varies in literature. In this study, w... more The definition of Luminal-B subclass of breast cancer (BC) varies in literature. In this study, we have compared the proliferation status; assessed using KI67 labeling index (KI67-LI), and HER2-expression in estrogen receptor positive (ER?) BC to assess their impact on the biological and clinical characteristics of luminal-BC. 1547 (73.8 %) well-characterized clinically annotated stage I-III ER ? BC were assessed for expression of KI67, HER2 (ASCO guidelines), and a large panel of relevant biomarkers (no = 37). 46.3 % of the cases show high KI67-LI ([13 %) and 8.4 % show HER2? and both markers are positively associated with younger age, higher tumor grade and poorer outcome. High KI67-LI and HER2? are associated with upregulation of ER-coactivators and proliferation-related markers and with downregulation of good prognostic markers. High KI67-LI is associated with larger size, advanced stage, and lymphovascular invasion (LVI) and with downregulation of luminal-enriched and DNA-damage repair markers. In contrast, HER2? is associated with upregulation of ERregulated proteins and E-cadherin. When analysis is restricted to high KI67-LI subgroup, HER2? shows an association with upregulation of differentiation-associated proteins and E-cadherin. Conversely, within HER2? class, high KI67-LI maintains its association with downregulation of differentiation-associated/luminal-enriched proteins. Outcome analyses indicate that both markers are independently associated with shorter survival but HER2? is associated with a worse outcome. Although both are associated with high proliferation and poor prognosis within ER ? BC, HER2? is less frequent than high KI67-LI. Unlike KI67, HER2 seems to independently drive the aggressive behavior of ER? tumors without downregulation of luminal proteins.
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Papers by Alaa Alshareeda