Dihydropyrimidinase-like 2 (DPYSL2) regulates breast cancer migration via a JAK/STAT3/vimentin axis

The intricate neuronal wiring during development requires cytoskeletal reorganization orchestrated by signaling cues. Considering that cytoskeletal remodeling is a hallmark of cell migration, we inquired whether metastatic cancer cells exploit the axon guidance proteins to migrate. Indeed, in breast cancer patients, we found a significant correlation between the mesenchymal markers and the expression of dihydropyrimidinase-like 2 (DPYSL2), a regulator of cytoskeletal dynamics in growing axons. Strikingly, DPYSL2 knockout in mesenchymal-like cells profoundly inhibited cell migration, invasion, stemness features, tumor growth rate, and metastasis. Next, we aimed to decode the molecular mechanism underlying this phenomenon and revealed an interaction between DPYSL2 and Janus kinase 1 (JAK1). This binding is crucial for triggering signal transducer and activator of transcription 3 (STAT3) and subsequently expressing vimentin, the pro-migratory intermediate filament. Collectively, we identified DPYSL2 as a molecular link between oncogenic signaling pathways and cytoskeletal reorganization in migrating breast cancer cells.

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Molecular and cellular mechanisms underlying brain metastasis of breast cancer

Cancer and Metastasis Reviews ◽

10.1007/s10555-020-09881-y ◽

2020 ◽

Vol 39 (3) ◽

pp. 711-720 ◽

Cited By ~ 2

Author(s):

Mari Hosonaga ◽

Hideyuki Saya ◽

Yoshimi Arima

Keyword(s):

Breast Cancer ◽

Brain Metastasis ◽

Brain Metastases ◽

Cancer Cells ◽

Metastatic Cancer ◽

Breast Cancer Patients ◽

Cellular Mechanisms ◽

Her2 Positive ◽

Cells Of The Microenvironment ◽

The Brain

Abstract Metastasis of cancer cells to the brain occurs frequently in patients with certain subtypes of breast cancer. In particular, patients with HER2-positive or triple-negative breast cancer are at high risk for the development of brain metastases. Despite recent advances in the treatment of primary breast tumors, the prognosis of breast cancer patients with brain metastases remains poor. A better understanding of the molecular and cellular mechanisms underlying brain metastasis might be expected to lead to improvements in the overall survival rate for these patients. Recent studies have revealed complex interactions between metastatic cancer cells and their microenvironment in the brain. Such interactions result in the activation of various signaling pathways related to metastasis in both cancer cells and cells of the microenvironment including astrocytes and microglia. In this review, we focus on such interactions and on their role both in the metastatic process and as potential targets for therapeutic intervention.

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Troglitazone Derivatives Induce Early Modifications of the Cytoskeleton and Inhibition of Migration in Breast Cancer Cells

10.21203/rs.3.rs-911353/v1 ◽

2021 ◽

Author(s):

Geoffroy Marine ◽

Lemesle Marine ◽

Kleinclauss Alexandra ◽

Mazerbourg Sabine ◽

Batista Levy ◽

...

Keyword(s):

Breast Cancer ◽

Cell Migration ◽

Cancer Cells ◽

Cell Lines ◽

Breast Cancer Cells ◽

Morphological Changes ◽

New Drugs ◽

Breast Cancer Cell Lines ◽

Breast Cancer Patients ◽

Hs578t Cell

Abstract Background The 5-years survival rate of breast cancer patients is around 90%. Unfortunately, some types of tumors, especially the triple negative breast cancer (TNBC), present chemo-resistance and have a higher relapse 5 years post-treatment due to metastasis. These challenges highlight the need for the development of new drugs for these tumors. In this context, we developed new troglitazone derivatives as support for such potential new treatment.Methods The kinetic of early cellular events was investigated after Δ2-TGZ and AB186 treatment by real-time cell analysis system (RTCA) in MCF-7 and MDA-MB-231 breast cancer cells, followed by cell morphology analysis by immuno-localization. Then, we characterized the action of both compounds on the cell migration by wound healing and transwell assays in TNBC MDA-MB-231 and Hs578T cell lines. Finally, we performed surface plasmon resonance (SPR) analysis and pull-down assay using biotinylated AB186 to identify cytoplasmic targets.Results Δ2-TGZ and AB186 induced a rapid modification of impedance-based signals and morphology in MCF7 and MDA-MB-231 breast cancer cell lines. This process was associated with an inhibition of cell migration in MDA-MB-231 and Hs578T cell lines. Subsequently, 6 cytoskeleton components have been identified as potential targets of AB186 in MDA-MB-231 cytoplasmic fraction. We further validated α-tubulin as one of the direct targets of AB186.Conclusion New troglitazone derivatives, Δ2-TGZ and AB186, induced early cell morphological changes and showed anti-migratory effects on TNBC cells suggesting that these drugs could be proposed as novel candidates to treat TNBC patients.

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Active Roles of Tumor Stroma in Breast Cancer Metastasis

International Journal of Breast Cancer ◽

10.1155/2012/574025 ◽

2012 ◽

Vol 2012 ◽

pp. 1-10 ◽

Cited By ~ 45

Author(s):

Zahraa I. Khamis ◽

Ziad J. Sahab ◽

Qing-Xiang Amy Sang

Keyword(s):

Breast Cancer ◽

Cancer Cells ◽

Tumor Cells ◽

Stromal Cells ◽

Cancer Metastasis ◽

Metastatic Cancer ◽

Breast Cancer Metastasis ◽

Secondary Site ◽

Signal Pathways ◽

Breast Cancer Patients

Metastasis is the major cause of death for breast cancer patients. Tumors are heterogenous cellular entities composed of cancer cells and cells of the microenvironment in which they reside. A reciprocal dynamic interaction occurs between the tumor cells and their surrounding stroma under physiological and pathological conditions. This tumor-host communication interface mediates the escape of tumor cells at the primary site, survival of circulating cancer cells in the vasculature, and growth of metastatic cancer at secondary site. Each step of the metastatic process is accompanied by recruitment of stromal cells from the microenvironment and production of unique array of growth factors and chemokines. Stromal microenvironment may play active roles in breast cancer metastasis. Elucidating the types of cells recruited and signal pathways involved in the crosstalk between tumor cells and stromal cells will help identify novel strategies for cotargeting cancer cells and tumor stromal cells to suppress metastasis and improve patient outcome.

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GPER1 is regulated by insulin in cancer cells and cancer-associated fibroblasts

Endocrine Related Cancer ◽

10.1530/erc-14-0245 ◽

2014 ◽

Vol 21 (5) ◽

pp. 739-753 ◽

Cited By ~ 30

Author(s):

Paola De Marco ◽

Enrica Romeo ◽

Adele Vivacqua ◽

Roberta Malaguarnera ◽

Sergio Abonante ◽

...

Keyword(s):

Breast Cancer ◽

Cancer Cells ◽

Cancer Progression ◽

Cell Cycle Progression ◽

Metastatic Cancer ◽

Promoter Sequence ◽

Serum Levels ◽

Estrogen Signaling ◽

Cancer Associated Fibroblasts ◽

Breast Cancer Patients

Elevated insulin levels have been associated with an increased cancer risk as well as with aggressive and metastatic cancer phenotypes characterized by a poor prognosis. Insulin stimulates the proliferation, migration, and invasiveness of cancer cells through diverse transduction pathways, including estrogen signaling. As G protein estrogen receptor 1 (GPER1) mediates rapid cell responses to estrogens, we evaluated the potential of insulin to regulate GPER1 expression and function in leiomyosarcoma cancer cells (SKUT-1) and breast cancer-associated fibroblasts (CAFs), which were used as a model system. We found that insulin transactivates the GPER1 promoter sequence and increases the mRNA and protein expression of GPER1 through the activation of the PRKCD/MAPK1/c-Fos/AP1 transduction pathway, as ascertained by means of specific pharmacological inhibitors and gene-silencing experiments. Moreover, cell migration triggered by insulin occurred through GPER1 and its main target gene CTGF, whereas the insulin-induced expression of GPER1 boosted cell-cycle progression and the glucose uptake stimulated by estrogens. Notably, a positive correlation between insulin serum levels and GPER1 expression was found in cancer fibroblasts obtained from breast cancer patients. Altogether, our data indicate that GPER1 may be included among the complex network of transduction signaling triggered by insulin that drives cells toward cancer progression.

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Inhibition of Ras and STAT3 activity of 4-(tert-butyl)-N-carbamoylbenzamide as antiproliferative agent in HER2-expressing breast cancer cells

Journal of Basic and Clinical Physiology and Pharmacology ◽

10.1515/jbcpp-2020-0508 ◽

2021 ◽

Vol 32 (4) ◽

pp. 363-371

Author(s):

Aguslina Kirtishanti ◽

Siswandono Siswodihardjo ◽

I Ketut Sudiana ◽

Desak G. A. Suprabawati ◽

Aristika Dinaryanti

Keyword(s):

Breast Cancer ◽

Monoclonal Antibodies ◽

Cancer Cells ◽

Cancer Patients ◽

Cancer Cell ◽

Breast Cancer Cells ◽

Janus Kinase ◽

Her2 Overexpression ◽

Breast Cancer Patients ◽

Her2 Signaling

Abstract Objectives Human epidermal growth factor receptor type 2 (HER2)-expressing breast cancer patients indicate poor prognosis in disease progression. HER2 overexpression can increase activities of Ras-mitogen activated protein kinase (Ras-MAPK) pathway and Janus Kinase (JAK)-STAT3, increasing breast cancer cell proliferation as demonstrated by marker Ki67. Therapeutic options for HER2-expressing breast cancer are limited and have major side effects, so anticancer development as an antiproliferative is needed. From previous research, synthetic chemical 4-(tert-butyl)-N-carbamoylbenzamide (4TBCB) compound has cytotoxic activity in vitro on HER2-expressing breast cancer cells. This study wanted to determine the mechanism 4TBCB compound in inhibiting HER2 signaling through Rat Sarcoma (Ras) and signal transducer and activator of transcription 3 (STAT3) pathway in HER2-expressing breast cancer cells. Methods Breast cancer cells were isolated from the biopsy tissue of breast cancer patients. The isolated cells were cultured and given 4TBCB test compound with three concentrations (0.305, 0.61, and 1.22 mM) and lapatinib 0.05 mM as a comparison compound. Cancer cell cultures were stained with monoclonal antibodies phosphorylated HER2 (pHER2), phosphorylated Ras (pRas), phosphorylated STAT3 (pSTAT3), and Ki67. The expression of pHER2, pRas, pSTAT3, and Ki67 proteins was observed using the immunofluorescence method and the results were compared with control cells, namely cancer cells that were not given 4TBCB and lapatinib but stained with monoclonal antibodies. Results 4TBCB compounds (0.61 and 1.22 mM) and lapatinib can reduce pHER2, pRas, pSTAT3, and Ki67 expressions compared to control cells. Conclusions 4TBCB compounds (0.61 and 1.22 mM) can reduce pHER2, pRas, pSTAT3, Ki67 expressions and predicted to inhibit HER2 signaling through the Ras and STAT3 pathways in HER2-expressing breast cancer cells.

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The Exposure of Breast Cancer Cells to Fulvestrant and Tamoxifen Modulates Cell Migration Differently

BioMed Research International ◽

10.1155/2013/147514 ◽

2013 ◽

Vol 2013 ◽

pp. 1-14 ◽

Cited By ~ 8

Author(s):

Dionysia Lymperatou ◽

Efstathia Giannopoulou ◽

Angelos K. Koutras ◽

Haralabos P. Kalofonos

Keyword(s):

Breast Cancer ◽

Cell Migration ◽

Cancer Cells ◽

Hormonal Therapy ◽

Breast Cancer Cells ◽

Protective Role ◽

Current Evidence ◽

Migration And Invasion ◽

Breast Cancer Patients ◽

Epithelial Tumor

There is no doubt that there are increased benefits of hormonal therapy to breast cancer patients; however, current evidence suggests that estrogen receptor (ER) blockage using antiestrogens is associated with a small induction of invasivenessin vitro. The mechanism by which epithelial tumor cells escape from the primary tumor and colonize to a distant site is not entirely understood. This study investigates the effect of two selective antagonists of the ER, Fulvestrant (Fulv) and Tamoxifen (Tam), on the invasive ability of breast cancer cells. We found that 17β-estradiol (E2) demonstrated a protective role regarding cell migration and invasion. Fulv did not alter this effect while Tam stimulated active cell migration according to an increase in Snail and a decrease in E-cadherin protein expression. Furthermore, both tested agents increased expression of matrix metalloproteinases (MMPs) and enhanced invasive potential of breast cancer cells. These changes were in line with focal adhesion kinase (FAK) rearrangement. Our data indicate that the anti-estrogens counteracted the protective role of E2concerning migration and invasion since their effect was not limited to antiproliferative events. Although Fulv caused a less aggressive result compared to Tam, the benefits of hormonal therapy concerning invasion and metastasis yet remain to be investigated.

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Targeting PKM2 promotes chemosensitivity of breast cancer cells in vitro and in vivo

Cancer Biomarkers ◽

10.3233/cbm-210111 ◽

2021 ◽

pp. 1-10

Author(s):

Yu Wang ◽

Han Zhao ◽

Ping Zhao ◽

Xingang Wang

Keyword(s):

Breast Cancer ◽

Neoadjuvant Chemotherapy ◽

Cancer Cells ◽

Cancer Patients ◽

Poor Prognosis ◽

Breast Cancer Cells ◽

Breast Cancer Patients ◽

Cancer Tissues

BACKGROUND: Pyruvate kinase M2 (PKM2) was overexpressed in many cancers, and high PKM2 expression was related with poor prognosis and chemoresistance. OBJECTIVE: We investigated the expression of PKM2 in breast cancer and analyzed the relation of PKM2 expression with chemotherapy resistance to the neoadjuvant chemotherapy (NAC). We also investigated whether PKM2 could reverse chemoresistance in breast cancer cells in vitro and in vivo. METHODS: Immunohistochemistry (IHC) was performed in 130 surgical resected breast cancer tissues. 78 core needle biopsies were collected from breast cancer patients before neoadjuvant chemotherapy. The relation of PKM2 expression and multi-drug resistance to NAC was compared. The effect of PKM2 silencing or overexpression on Doxorubicin (DOX) sensitivity in the MCF-7 cells in vitro and in vivo was compared. RESULTS: PKM2 was intensively expressed in breast cancer tissues compared to adjacent normal tissues. In addition, high expression of PKM2 was associated with poor prognosis in breast cancer patients. The NAC patients with high PKM2 expression had short survival. PKM2 was an independent prognostic predictor for surgical resected breast cancer and NAC patients. High PKM2 expression was correlated with neoadjuvant treatment resistance. High PKM2 expression significantly distinguished chemoresistant patients from chemosensitive patients. In vitro and in vivo knockdown of PKM2 expression decreases the resistance to DOX in breast cancer cells in vitro and tumors in vivo. CONCLUSION: PKM2 expression was associated with chemoresistance of breast cancers, and could be used to predict the chemosensitivity. Furthermore, targeting PKM2 could reverse chemoresistance, which provides an effective treatment methods for patients with breast cancer.

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The Mechanical Fingerprint of Circulating Tumor Cells (CTCs) in Breast Cancer Patients

Cancers ◽

10.3390/cancers13051119 ◽

2021 ◽

Vol 13 (5) ◽

pp. 1119

Author(s):

Ivonne Nel ◽

Erik W. Morawetz ◽

Dimitrij Tschodu ◽

Josef A. Käs ◽

Bahriye Aktas

Keyword(s):

Breast Cancer ◽

Cancer Cells ◽

Cancer Patients ◽

Tumor Cells ◽

Circulating Tumor Cells ◽

Hematopoietic Cells ◽

Surrogate Marker ◽

Clinical Samples ◽

Breast Cancer Patients ◽

Shape Restoration

Circulating tumor cells (CTCs) are a potential predictive surrogate marker for disease monitoring. Due to the sparse knowledge about their phenotype and its changes during cancer progression and treatment response, CTC isolation remains challenging. Here we focused on the mechanical characterization of circulating non-hematopoietic cells from breast cancer patients to evaluate its utility for CTC detection. For proof of premise, we used healthy peripheral blood mononuclear cells (PBMCs), human MDA-MB 231 breast cancer cells and human HL-60 leukemia cells to create a CTC model system. For translational experiments CD45 negative cells—possible CTCs—were isolated from blood samples of patients with mamma carcinoma. Cells were mechanically characterized in the optical stretcher (OS). Active and passive cell mechanical data were related with physiological descriptors by a random forest (RF) classifier to identify cell type specific properties. Cancer cells were well distinguishable from PBMC in cell line tests. Analysis of clinical samples revealed that in PBMC the elliptic deformation was significantly increased compared to non-hematopoietic cells. Interestingly, non-hematopoietic cells showed significantly higher shape restoration. Based on Kelvin–Voigt modeling, the RF algorithm revealed that elliptic deformation and shape restoration were crucial parameters and that the OS discriminated non-hematopoietic cells from PBMC with an accuracy of 0.69, a sensitivity of 0.74, and specificity of 0.63. The CD45 negative cell population in the blood of breast cancer patients is mechanically distinguishable from healthy PBMC. Together with cell morphology, the mechanical fingerprint might be an appropriate tool for marker-free CTC detection.

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Proteomic Analysis of Zeb1 Interactome in Breast Carcinoma Cells

Molecules ◽

10.3390/molecules26113143 ◽

2021 ◽

Vol 26 (11) ◽

pp. 3143

Author(s):

Sergey E. Parfenyev ◽

Sergey V. Shabelnikov ◽

Danila Y. Pozdnyakov ◽

Olga O. Gnedina ◽

Leonid S. Adonin ◽

...

Keyword(s):

Breast Cancer ◽

Cancer Cells ◽

Epithelial To Mesenchymal Transition ◽

Critical Role ◽

Malignant Neoplasm ◽

Malignant Neoplasms ◽

Breast Cancer Patients ◽

Survival Prognosis ◽

Mesenchymal Transition ◽

Wide Range

Breast cancer is the most frequently diagnosed malignant neoplasm and the second leading cause of cancer death among women. Epithelial-to-mesenchymal Transition (EMT) plays a critical role in the organism development, providing cell migration and tissue formation. However, its erroneous activation in malignancies can serve as the basis for the dissemination of cancer cells and metastasis. The Zeb1 transcription factor, which regulates the EMT activation, has been shown to play an essential role in malignant transformation. This factor is involved in many signaling pathways that influence a wide range of cellular functions via interacting with many proteins that affect its transcriptional functions. Importantly, the interactome of Zeb1 depends on the cellular context. Here, using the inducible expression of Zeb1 in epithelial breast cancer cells, we identified a substantial list of novel potential Zeb1 interaction partners, including proteins involved in the formation of malignant neoplasms, such as ATP-dependent RNA helicase DDX17and a component of the NURD repressor complex, CTBP2. We confirmed the presence of the selected interactors by immunoblotting with specific antibodies. Further, we demonstrated that co-expression of Zeb1 and CTBP2 in breast cancer patients correlated with the poor survival prognosis, thus signifying the functionality of the Zeb1–CTBP2 interaction.

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Early, On-Treatment Levels and Dynamic Changes of Genomic Instability in Circulating Tumor DNA Predict Response to Treatment and Outcome in Metastatic Breast Cancer Patients

Cancers ◽

10.3390/cancers13061331 ◽

2021 ◽

Vol 13 (6) ◽

pp. 1331

Author(s):

Adriana Aguilar-Mahecha ◽

Josiane Lafleur ◽

Susie Brousse ◽

Olga Savichtcheva ◽

Kimberly A. Holden ◽

...

Keyword(s):

Breast Cancer ◽

Metastatic Breast Cancer ◽

Genomic Instability ◽

Metastatic Cancer ◽

Metastatic Breast ◽

Circulating Tumor Dna ◽

Response To Treatment ◽

Breast Cancer Patients ◽

T1 And T2 ◽

Tumor Dna

Background: Circulating tumor DNA (ctDNA) offers high sensitivity and specificity in metastatic cancer. However, many ctDNA assays rely on specific mutations in recurrent genes or require the sequencing of tumor tissue, difficult to do in a metastatic disease. The purpose of this study was to define the predictive and prognostic values of the whole-genome sequencing (WGS) of ctDNA in metastatic breast cancer (MBC). Methods: Plasma from 25 patients with MBC were taken at the baseline, prior to treatment (T0), one week (T1) and two weeks (T2) after treatment initiation and subjected to low-pass WGS. DNA copy number changes were used to calculate a Genomic Instability Number (GIN). A minimum predefined GIN value of 170 indicated detectable ctDNA. GIN values were correlated with the treatment response at three and six months by Response Evaluation Criteria in Solid Tumours assessed by imaging (RECIST) criteria and with overall survival (OS). Results: GIN values were detectable (>170) in 64% of patients at the baseline and were significantly prognostic (41 vs. 18 months OS for nondetectable vs. detectable GIN). Detectable GIN values at T1 and T2 were significantly associated with poor OS. Declines in GIN at T1 and T2 of > 50% compared to the baseline were associated with three-month response and, in the case of T1, with OS. On the other hand, a rise in GIN at T2 was associated with a poor response at three months. Conclusions: Very early measurements using WGS of cell-free DNA (cfDNA) from the plasma of MBC patients provided a tumor biopsy-free approach to ctDNA measurement that was both predictive of the early tumor response at three months and prognostic.

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