The natural hormone 17β-estradiol (17β-E2) is known to induce tumor angiogenesis in various targe... more The natural hormone 17β-estradiol (17β-E2) is known to induce tumor angiogenesis in various target organs by activating positive regulators of angiogenesis. In this study, we show for the first time that in human umbilical vein endothelial cells (HUVECs), 17β-E2 transiently down-regulates the expression and secretion of a potent negative regulator of angiogenesis, thrombospondin-1 (TSP-1). This inhibitory effect of 17β-E2 is mediated through nongenomic estrogen receptor (ER)/mitogen-activated protein kinase (MAPK)/extracellular-regulated kinase (ERK) 1/2 and c-Jun NH2-terminal kinase (JNK)/stress-activated protein kinase (SAPK) signaling pathways, because this effect can be abolished by a pure ER antagonist (ICI 182,780) and inhibitors of downstream signaling proteins of MAPK signaling cascades, including MAPK kinase 1/2 and ERK1/2 inhibitor and JNK/SAPK inhibitor. To understand the functional role(s) of TSP-1 during estradiol-induced angiogenesis, we examined the growth and migrati...
Estrogens have been shown to regulate vascular endothelial growth factor-A (VEGF-A) for physiolog... more Estrogens have been shown to regulate vascular endothelial growth factor-A (VEGF-A) for physiological and patho-physiological functions. However, estrogen action on VEGF-A mRNA expression has not been completely elucidated. We have identified two phases of activation of VEGF-A mRNA transcription, one early and one late response, induced by 17beta-estradiol (17beta-E2) in ER+ MCF-7 breast tumor cells, depending upon the length of exposure. VEGF-A mRNA level was significantly higher than control in tumor cells after 2 h of 17beta-E2 exposure. Furthermore this induction was not inhibited by cycloheximide, indicating that it was a direct effect of estrogen. In contrast VEGF-A mRNA expression was back at basal level in MCF-7 cells exposed to 17beta-E2 for 6 h. However, expression levels were again significantly augmented after 24 h of exposure, and this induction was unaltered by cycloheximide indicating that de novo protein synthesis was not required and like early response, it was a di...
Non-small-cell lung cancer (NSCLC) patients with epidermal growth factor receptor (EGFR) mutation... more Non-small-cell lung cancer (NSCLC) patients with epidermal growth factor receptor (EGFR) mutation eventually develop resistance to EGFR-targeted tyrosine kinase inhibitors (TKIs). Treatment resistance remains the primary obstacle to the successful treatment of NSCLC. Although drug resistance mechanisms have been studied extensively in NSCLC, the regulation of these mechanisms has not been completely understood. Recently, increasing numbers of microRNAs (miRNAs) are implicated in EGFR-TKI resistance, indicating that miRNAs may serve as novel targets and may hold promise as predictive biomarkers for anti-EGFR therapy. MicroRNA-506 (miR-506) has been identified as a tumor suppressor in many cancers, including lung cancer; however, the role of miR-506 in lung cancer chemoresistance has not yet been addressed. Here we report that miR-506-3p expression was markedly reduced in erlotinib-resistant (ER) cells. We identified Sonic Hedgehog (SHH) as a novel target of miR-506-3p, aberrantly act...
Mouse models of breast cancer with specific molecular subtypes (e.g., ER or HER2 positive) in an ... more Mouse models of breast cancer with specific molecular subtypes (e.g., ER or HER2 positive) in an immunocompetent or an immunocompromised environment significantly contribute to our understanding of cancer biology, despite some limitations, and they give insight into targeted therapies. However, an ideal triple-negative breast cancer (TNBC) mouse model is lacking. What has been missing in the TNBC mouse model is a sequential progression of the disease in an essential native microenvironment. This notion inspired us to develop a TNBC-model in syngeneic mice using a mammary intraductal (MIND) method. To achieve this goal, Mvt-1and 4T1 TNBC mouse cell lines were injected into the mammary ducts via nipples of FVB/N mice and BALB/c wild-type immunocompetent mice, respectively. We established that the TNBC-MIND model in syngeneic mice could epitomize all breast cancer progression stages and metastasis into the lungs via lymphatic or hematogenous dissemination within four weeks. Collectivel...
Journal of cell communication and signaling, Jan 29, 2017
Breast cancer (BC) has emerged as a deadly disease that affects the lives of millions of women wo... more Breast cancer (BC) has emerged as a deadly disease that affects the lives of millions of women worldwide. It is the second leading cause of cancer-related deaths in the United States. Advancements in BC screening, preventive measures and treatment have resulted in significant decline in BC related deaths. However, unacceptable levels of racial disparity have been consistently reported, especially in African-American (AA) women compared to European American (EA). AA women go through worse prognosis, shorter survival time and higher mortality rates, despite higher cancer incidence reported in EA. These disparities are independent of socioeconomic status, access to healthcare or age, or even the stage of BC. Recent race-specific genetic and epigenetic studies have reported biological causes, which form the crux of this review. However, the developments are just the tip of the iceberg. Prioritizing primary research towards studying race-specific tumor microenvironment and biological com...
Myc-associated zinc-finger protein (MAZ) is a transcription factor with dual roles in transcripti... more Myc-associated zinc-finger protein (MAZ) is a transcription factor with dual roles in transcription initiation and termination. Deregulation of MAZ expression is associated with the progression of pancreatic ductal adenocarcinoma (PDAC). However, the mechanism of action of MAZ in PDAC progression is largely unknown. Here, we present evidence that MAZ mRNA expression and protein levels are increased in human PDAC cell lines, tissue samples, a subcutaneous tumor xenograft in a nude mouse model, and spontaneous cancer in the genetically engineered PDAC mouse model. We also found that MAZ is predominantly expressed in pancreatic cancer stem cells. Functional analysis indicated that MAZ depletion in PDAC cells inhibits invasive phenotypes such as the epithelial-to-mesenchymal transition, migration, invasion, and the sphere-forming ability of PDAC cells. Mechanistically, we detected no direct effects of MAZ on the expression of K-Ras mutants, but MAZ increased the activity of CRAF-ERK sig...
Breast cancer progression and relapse is conceivably due to tumor initiating cells (TICs)/cancer ... more Breast cancer progression and relapse is conceivably due to tumor initiating cells (TICs)/cancer stem cells. EMT (epithelial-mesenchymal-transition)-signaling regulates TICs' turnover. However, the mechanisms associated with this episode are unclear. We show that, in triple-negative-breast cancer (TNBC) cells enriched with TICs, CCN5 significantly blocks cellular growth via apoptosis, reversing EMT-signaling and impairing mammosphere formation, thereby blocking the tumor-forming ability and invasive capacity of these cells. To corroborate these findings, we isolated tumor-initiating side populations (SP) and non-side population (NSP or main population) from MCF-7 cell line, and evaluated the impact of CCN5 on these subpopulations. CCN5 was overexpressed in the NSP but downregulated in the SP. Characteristically, NSP cells are ER-α positive and epithelial type with little tumorigenic potency, while SP cells are very similar to triple-negative ones that do not express ER-α- and He...
Pancreatic Ductal Adenocarcinoma (PDAC) is among the most aggressive of solid malignancies respon... more Pancreatic Ductal Adenocarcinoma (PDAC) is among the most aggressive of solid malignancies responsible for around 330,000 deaths globally and accounts for the fourth most common cause of deaths due to cancer in USA. There has been a little advancement in the treatment of PDAC patients and the mortality rate has remained unchanged and may even be climbing up. Metformin, an oral biguanide medication used to treat type-2 diabetes mellitus, has demonstrated potential therapeutic effect against PDAC. Recent meta-analysis and epidemiologic studies indicate that diabetic patients treated with metformin were less likely to develop pancreatic cancer exhibit longer overall survival than those using other oral antidiabetic medications. Although there are several reports of the possible mode of action of Metformin against PDAC, no direct cellular/ molecular target is till reported. The present study aims to identify the direct molecular target of metformin in pancreatic cancer cells. Previously, we have demonstrated that matricellular protein CCN1/ Cyr61 plays pivotal role in pancreatic cancer development, maintenance of stemness, and induction of tumor angiogenesis. Hence the goal of this study is to investigate whether CCN1/ Cyr61 signaling cascade acts as a potential target for metformin. The encouraging results we have obtained so far reveal that metformin, which reduces CCN1 expression, significantly inhibits SDF-1 induced invasion, formation of tumor spheres (pancospheres) and also results in the down regulation of CXCR4 receptor in PC cell lines (Panc-1 and AsPC-1). Interestingly we observed that CXCR4 expression is drastically down regulated in genetically engineered CCN1-knock out Panc-1 (Panc-1 KO CCN1 ) cells, compared to the scrambled shRNA transfected Panc-1 cells. Further, it is also observed that Panc-1 K. CCN1 cells are found to be more susceptible to metformin treatment, whereas extracellular supplementation of recombinant CCN1 protein significantly abrogates the effect of metformin on PC cells. Moreover, tumor progression was found to be drastically inhibited in metformin treated Panc-1 (CCN1+ve) tumor xenografts in nude mice model, while the anti-tumorigenic effect of metformin was found to be more drastic in Panc-1 KO CCN1 tumor xenografts. In this scenario, we conclude that CCN1 acts as a direct target for metformin in PC cells, and targeted knock down of CCN1 increases the effectiveness of metformin. [This project is funded by VA Merit Award grants (SB & SKB)] Citation Format: Amlan Das, Archana De, Inamul Haque, Gargi Maity, Sushanta Banerjee, Snigdha Banerjee. Metformin inhibits the oncogenic potential and invasiveness of pancreatic cancer cells targeting CCN1-CXCR4 axis : A new perspective for an old antidiabetic drug. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3572. doi:10.1158/1538-7445.AM2015-3572
Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA CCN5, a matricellular prote... more Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA CCN5, a matricellular protein, represents promising treatment target in triple negative breast cancer (TNBC) as treatment or induced activation of CCN5 in TNBC cells promotes cell growth arrest at G0/G1 phase, reduces cell proliferation and delays tumor growth in xenograft model. Our studies found that p27Kip1 tumor suppressor protein is upregulated and relocalized to the nucleus from cytoplasm by CCN5 in these cells and that these two events (i.e., upregulation and relocalization) of p27Kip1 is required for CCN5-induced growth inhibition of TNBC cells. In the absence of CCN5, p27Kip1 remains mostly in the cytoplasm, a state of aggressive nature of cancer cells. Mechanistically, CCN5 inhibits Skp2 expression, which seems to stabilize p27Kip1 protein in these cells. On the other hand, CCN5 also recruits FOXO3a to mediate transcriptional regulation of p27Kip1. The recruitment of FOXO3a is achieved by the induction of its expression and activity through shifting from cytoplasm to the nucleus. Our data indicate that CCN5 blocks PI3K/AKT signaling to dephosphorylate at S318, S253 and Thr32 in FOXO3a for nuclear relocalization and activation of FOXO3a. Inhibition of α6β1 diminishes CCN5 action on p27Kip1 in TNBC cells. Collectively, CCN5 may have therapeutic potential for TNBC. Citation Format: Inamul Haque, Snigdha Banerjee, Archana De, Gargi Maity, Sandipto Sarkar, Douglas McGragor, Sushanta K. Banerjee. CCN5/wisp-2 induced growth arrest of aggressive triple negative breast cancer cells is promoted through accumulation and trafficking of p27kip1 . [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1552. doi:10.1158/1538-7445.AM2014-1552
The natural hormone 17β-estradiol (17β-E2) is known to induce tumor angiogenesis in various targe... more The natural hormone 17β-estradiol (17β-E2) is known to induce tumor angiogenesis in various target organs by activating positive regulators of angiogenesis. In this study, we show for the first time that in human umbilical vein endothelial cells (HUVECs), 17β-E2 transiently down-regulates the expression and secretion of a potent negative regulator of angiogenesis, thrombospondin-1 (TSP-1). This inhibitory effect of 17β-E2 is mediated through nongenomic estrogen receptor (ER)/mitogen-activated protein kinase (MAPK)/extracellular-regulated kinase (ERK) 1/2 and c-Jun NH2-terminal kinase (JNK)/stress-activated protein kinase (SAPK) signaling pathways, because this effect can be abolished by a pure ER antagonist (ICI 182,780) and inhibitors of downstream signaling proteins of MAPK signaling cascades, including MAPK kinase 1/2 and ERK1/2 inhibitor and JNK/SAPK inhibitor. To understand the functional role(s) of TSP-1 during estradiol-induced angiogenesis, we examined the growth and migrati...
Estrogens have been shown to regulate vascular endothelial growth factor-A (VEGF-A) for physiolog... more Estrogens have been shown to regulate vascular endothelial growth factor-A (VEGF-A) for physiological and patho-physiological functions. However, estrogen action on VEGF-A mRNA expression has not been completely elucidated. We have identified two phases of activation of VEGF-A mRNA transcription, one early and one late response, induced by 17beta-estradiol (17beta-E2) in ER+ MCF-7 breast tumor cells, depending upon the length of exposure. VEGF-A mRNA level was significantly higher than control in tumor cells after 2 h of 17beta-E2 exposure. Furthermore this induction was not inhibited by cycloheximide, indicating that it was a direct effect of estrogen. In contrast VEGF-A mRNA expression was back at basal level in MCF-7 cells exposed to 17beta-E2 for 6 h. However, expression levels were again significantly augmented after 24 h of exposure, and this induction was unaltered by cycloheximide indicating that de novo protein synthesis was not required and like early response, it was a di...
Non-small-cell lung cancer (NSCLC) patients with epidermal growth factor receptor (EGFR) mutation... more Non-small-cell lung cancer (NSCLC) patients with epidermal growth factor receptor (EGFR) mutation eventually develop resistance to EGFR-targeted tyrosine kinase inhibitors (TKIs). Treatment resistance remains the primary obstacle to the successful treatment of NSCLC. Although drug resistance mechanisms have been studied extensively in NSCLC, the regulation of these mechanisms has not been completely understood. Recently, increasing numbers of microRNAs (miRNAs) are implicated in EGFR-TKI resistance, indicating that miRNAs may serve as novel targets and may hold promise as predictive biomarkers for anti-EGFR therapy. MicroRNA-506 (miR-506) has been identified as a tumor suppressor in many cancers, including lung cancer; however, the role of miR-506 in lung cancer chemoresistance has not yet been addressed. Here we report that miR-506-3p expression was markedly reduced in erlotinib-resistant (ER) cells. We identified Sonic Hedgehog (SHH) as a novel target of miR-506-3p, aberrantly act...
Mouse models of breast cancer with specific molecular subtypes (e.g., ER or HER2 positive) in an ... more Mouse models of breast cancer with specific molecular subtypes (e.g., ER or HER2 positive) in an immunocompetent or an immunocompromised environment significantly contribute to our understanding of cancer biology, despite some limitations, and they give insight into targeted therapies. However, an ideal triple-negative breast cancer (TNBC) mouse model is lacking. What has been missing in the TNBC mouse model is a sequential progression of the disease in an essential native microenvironment. This notion inspired us to develop a TNBC-model in syngeneic mice using a mammary intraductal (MIND) method. To achieve this goal, Mvt-1and 4T1 TNBC mouse cell lines were injected into the mammary ducts via nipples of FVB/N mice and BALB/c wild-type immunocompetent mice, respectively. We established that the TNBC-MIND model in syngeneic mice could epitomize all breast cancer progression stages and metastasis into the lungs via lymphatic or hematogenous dissemination within four weeks. Collectivel...
Journal of cell communication and signaling, Jan 29, 2017
Breast cancer (BC) has emerged as a deadly disease that affects the lives of millions of women wo... more Breast cancer (BC) has emerged as a deadly disease that affects the lives of millions of women worldwide. It is the second leading cause of cancer-related deaths in the United States. Advancements in BC screening, preventive measures and treatment have resulted in significant decline in BC related deaths. However, unacceptable levels of racial disparity have been consistently reported, especially in African-American (AA) women compared to European American (EA). AA women go through worse prognosis, shorter survival time and higher mortality rates, despite higher cancer incidence reported in EA. These disparities are independent of socioeconomic status, access to healthcare or age, or even the stage of BC. Recent race-specific genetic and epigenetic studies have reported biological causes, which form the crux of this review. However, the developments are just the tip of the iceberg. Prioritizing primary research towards studying race-specific tumor microenvironment and biological com...
Myc-associated zinc-finger protein (MAZ) is a transcription factor with dual roles in transcripti... more Myc-associated zinc-finger protein (MAZ) is a transcription factor with dual roles in transcription initiation and termination. Deregulation of MAZ expression is associated with the progression of pancreatic ductal adenocarcinoma (PDAC). However, the mechanism of action of MAZ in PDAC progression is largely unknown. Here, we present evidence that MAZ mRNA expression and protein levels are increased in human PDAC cell lines, tissue samples, a subcutaneous tumor xenograft in a nude mouse model, and spontaneous cancer in the genetically engineered PDAC mouse model. We also found that MAZ is predominantly expressed in pancreatic cancer stem cells. Functional analysis indicated that MAZ depletion in PDAC cells inhibits invasive phenotypes such as the epithelial-to-mesenchymal transition, migration, invasion, and the sphere-forming ability of PDAC cells. Mechanistically, we detected no direct effects of MAZ on the expression of K-Ras mutants, but MAZ increased the activity of CRAF-ERK sig...
Breast cancer progression and relapse is conceivably due to tumor initiating cells (TICs)/cancer ... more Breast cancer progression and relapse is conceivably due to tumor initiating cells (TICs)/cancer stem cells. EMT (epithelial-mesenchymal-transition)-signaling regulates TICs' turnover. However, the mechanisms associated with this episode are unclear. We show that, in triple-negative-breast cancer (TNBC) cells enriched with TICs, CCN5 significantly blocks cellular growth via apoptosis, reversing EMT-signaling and impairing mammosphere formation, thereby blocking the tumor-forming ability and invasive capacity of these cells. To corroborate these findings, we isolated tumor-initiating side populations (SP) and non-side population (NSP or main population) from MCF-7 cell line, and evaluated the impact of CCN5 on these subpopulations. CCN5 was overexpressed in the NSP but downregulated in the SP. Characteristically, NSP cells are ER-α positive and epithelial type with little tumorigenic potency, while SP cells are very similar to triple-negative ones that do not express ER-α- and He...
Pancreatic Ductal Adenocarcinoma (PDAC) is among the most aggressive of solid malignancies respon... more Pancreatic Ductal Adenocarcinoma (PDAC) is among the most aggressive of solid malignancies responsible for around 330,000 deaths globally and accounts for the fourth most common cause of deaths due to cancer in USA. There has been a little advancement in the treatment of PDAC patients and the mortality rate has remained unchanged and may even be climbing up. Metformin, an oral biguanide medication used to treat type-2 diabetes mellitus, has demonstrated potential therapeutic effect against PDAC. Recent meta-analysis and epidemiologic studies indicate that diabetic patients treated with metformin were less likely to develop pancreatic cancer exhibit longer overall survival than those using other oral antidiabetic medications. Although there are several reports of the possible mode of action of Metformin against PDAC, no direct cellular/ molecular target is till reported. The present study aims to identify the direct molecular target of metformin in pancreatic cancer cells. Previously, we have demonstrated that matricellular protein CCN1/ Cyr61 plays pivotal role in pancreatic cancer development, maintenance of stemness, and induction of tumor angiogenesis. Hence the goal of this study is to investigate whether CCN1/ Cyr61 signaling cascade acts as a potential target for metformin. The encouraging results we have obtained so far reveal that metformin, which reduces CCN1 expression, significantly inhibits SDF-1 induced invasion, formation of tumor spheres (pancospheres) and also results in the down regulation of CXCR4 receptor in PC cell lines (Panc-1 and AsPC-1). Interestingly we observed that CXCR4 expression is drastically down regulated in genetically engineered CCN1-knock out Panc-1 (Panc-1 KO CCN1 ) cells, compared to the scrambled shRNA transfected Panc-1 cells. Further, it is also observed that Panc-1 K. CCN1 cells are found to be more susceptible to metformin treatment, whereas extracellular supplementation of recombinant CCN1 protein significantly abrogates the effect of metformin on PC cells. Moreover, tumor progression was found to be drastically inhibited in metformin treated Panc-1 (CCN1+ve) tumor xenografts in nude mice model, while the anti-tumorigenic effect of metformin was found to be more drastic in Panc-1 KO CCN1 tumor xenografts. In this scenario, we conclude that CCN1 acts as a direct target for metformin in PC cells, and targeted knock down of CCN1 increases the effectiveness of metformin. [This project is funded by VA Merit Award grants (SB & SKB)] Citation Format: Amlan Das, Archana De, Inamul Haque, Gargi Maity, Sushanta Banerjee, Snigdha Banerjee. Metformin inhibits the oncogenic potential and invasiveness of pancreatic cancer cells targeting CCN1-CXCR4 axis : A new perspective for an old antidiabetic drug. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3572. doi:10.1158/1538-7445.AM2015-3572
Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA CCN5, a matricellular prote... more Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA CCN5, a matricellular protein, represents promising treatment target in triple negative breast cancer (TNBC) as treatment or induced activation of CCN5 in TNBC cells promotes cell growth arrest at G0/G1 phase, reduces cell proliferation and delays tumor growth in xenograft model. Our studies found that p27Kip1 tumor suppressor protein is upregulated and relocalized to the nucleus from cytoplasm by CCN5 in these cells and that these two events (i.e., upregulation and relocalization) of p27Kip1 is required for CCN5-induced growth inhibition of TNBC cells. In the absence of CCN5, p27Kip1 remains mostly in the cytoplasm, a state of aggressive nature of cancer cells. Mechanistically, CCN5 inhibits Skp2 expression, which seems to stabilize p27Kip1 protein in these cells. On the other hand, CCN5 also recruits FOXO3a to mediate transcriptional regulation of p27Kip1. The recruitment of FOXO3a is achieved by the induction of its expression and activity through shifting from cytoplasm to the nucleus. Our data indicate that CCN5 blocks PI3K/AKT signaling to dephosphorylate at S318, S253 and Thr32 in FOXO3a for nuclear relocalization and activation of FOXO3a. Inhibition of α6β1 diminishes CCN5 action on p27Kip1 in TNBC cells. Collectively, CCN5 may have therapeutic potential for TNBC. Citation Format: Inamul Haque, Snigdha Banerjee, Archana De, Gargi Maity, Sandipto Sarkar, Douglas McGragor, Sushanta K. Banerjee. CCN5/wisp-2 induced growth arrest of aggressive triple negative breast cancer cells is promoted through accumulation and trafficking of p27kip1 . [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1552. doi:10.1158/1538-7445.AM2014-1552
Uploads