Inhibition of Cytokine Receptor gp130 Signaling in Breast Cancer Cells Markedly Suppresses Secretion of Tissue Factor-Expressing Tumor Microparticles.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 1935-1935
Author(s):  
Diego A. de Idiaquez ◽  
Dannielle E. Branam ◽  
Larry Lamb ◽  
Dongquan Chen ◽  
Lihong Teng ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Flow Cytometry ◽  
Cancer Cells ◽  
Tissue Factor ◽  
Tumor Invasion ◽  
Breast Cancer Cells ◽  
Molecular Mechanisms ◽  
Extracellular Proteins ◽  
Dominant Negative

Abstract gp130 is the common signaling receptor subunit for the IL-6 family of cytokines. We have previously shown that expression of a dominant negative (DN) gp130 inhibitory protein in MDA-MB-231 breast cancer cells decreased tumor invasion and angiogenesis in an orthotopic animal model (Cancer Research64:6924, 2004). In order to better understand the molecular mechanisms of this decreased malignancy, we determined global MDA-MB-231 gene expression in the presence of DN gp130. RNA was obtained from multiple independent single cell clones of MDA-MB-231 cells expressing either DN gp130 (n=5) or vector-only (n=5). Each clone was studied using Affymetrix U133A chips containing >22,000 genes. We identified 237 genes that were up- or down-regulated in the presence of DN gp130 at least 2-fold with p<0.05. Genes of interest were further studied by real-time PCR, western blot, and flow cytometry. We focused on extracellular proteins involved in proteolytic pathways. Six coagulation regulatory proteins were found to be differentially regulated by DN gp130: tissue factor (decreased), PAI-1 (decreased), alpha-2 antiplasmin (decreased), TFPI (increased), thrombomodulin (increased), and MMP-14 (increased). Thus, inhibition of gp130 signaling results in an anticoagulant phenotype in these cells. Tissue factor regulation was further studied. Cell surface tissue factor protein was down-regulated 10-fold in MDA-MB-231 DN gp130 cells compared to MDA-MB-231 control cells. Tumor microparticles (MP) were isolated from conditioned media by differential centrifugation. MDA-MB-231 MP contained uPA and MMP-9, as previously reported for MP from other tumors. Tissue factor was strongly expressed in control MP and nearly absent in MP from DN gp130 cells. Flow cytometry of tumor MP was used to quantitate tissue factor-expressing MP. There was a 50-fold decrease in double-labeled annexin V positive/tissue factor positive MP obtained from DN gp130 cells compared to control cells. In addition, the absolute number of tumor MP was decreased by 50% in the presence of DN gp130. In order to determine the mechanism of this regulation of breast cancer MP by gp130, the above list of 237 differentially expressed genes was re-examined. Proteins known to be enriched in MP- CD55, CD59, and three members of the tetraspanin family (including CD9) were significantly decreased in the presence of DN gp130. The expression of three proteins that are involved in the cellular production of MP were also significantly affected by DN gp130: Ca2+-dependent activator of protein for secretion 2, Rab11 interacting protein, and syntaxin 3. Our data supports a hypothesis in which signaling via IL-6 family cytokines induces a procoagulant gene expression program in breast cancer cells. As part of this program, the production of tumor MP are increased via gene regulation of cellular proteins that control MP processing. Additionally, the procoagulant properties of the tumor MP are increased due to an increase in tissue factor content. It has been reported that both activation of coagulation and production of tumor MP are involved in solid tumor invasion and metastasis. Inflammatory signaling by IL-6 family members in the tumor milieu could be important in this process. This hypothesis has clinical significance since potential pharmacologic approaches to specific gp130 inhibition are available.

scholarly journals LIN28A modulates splicing and gene expression programs in breast cancer cells

2015 ◽  
pp. MCB.00426-15 ◽  
Author(s):  
Jun Yang ◽  
Brian D. Bennett ◽  
Shujun Luo ◽  
Kaoru Inoue ◽  
Sara A. Grimm ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Cancer Cells ◽  
Cancer Biology ◽  
Breast Cancer Cells ◽  
Molecular Mechanisms ◽  
Rna Binding ◽  
Breast Cancer Subtype ◽  
Computational Method ◽  
Hnrnp A1

LIN28 is an evolutionarily conserved RNA-binding protein with critical functions in developmental timing and cancer. However molecular mechanisms underlying LIN28's oncogenic properties are yet to be described. RIP-Seq analysis revealed significant LIN28 binding within 843 mRNAs in breast cancer cells. Many of the LIN28 bound mRNAs are implicated in the regulation of RNA and cell metabolism. We identify heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1), a protein with multiple roles in mRNA metabolism, as a LIN28 interacting partner. Subsequently, we use a custom computational method to identify differentially spliced gene isoforms in LIN28 and hnRNP A1 siRNA-treated cells. Results reveal these proteins regulate alternative splicing and steady state mRNA expression of genes implicated in aspects of breast cancer biology. Notably, cells lacking LIN28 undergo significant isoform switching of the ENAH gene, resulting in a decrease in the expression of ENAH exon 11a isoform. Expression of ENAH isoform 11a has been shown to be elevated in breast cancers that express HER2. Intriguingly, analysis of publicly available TCGA array data reveals LIN28 expression is significantly different in HER2 compared to other breast cancer subtypes. Collectively, our data suggests that LIN28 may regulate splicing and gene expression programs that drive breast cancer subtype phenotypes.

scholarly journals The histone variant macroH2A1.1 regulates gene expression by direct association with their transcription start site

2020 ◽  
Author(s):  
Ludmila Recoules ◽  
Alexandre Heurteau ◽  
Flavien Raynal ◽  
Fatima Moutahir ◽  
Fabienne Bejjani ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Cancer Cells ◽  
Transcription Start Site ◽  
Breast Cancer Cells ◽  
Molecular Mechanisms ◽  
Histone Variants ◽  
Histone Variant ◽  
Start Site ◽  
Transcription Start

AbstractThe histone variant macroH2A1 (mH2A1) is involved in cellular growth, differentiation and reprogramming, but the underlying molecular mechanisms are a matter of debate. Different roles of mH2A1 in gene expression may relate to functional differences of its two splicing isoforms, mH2A1.1 and mH2A1.2. Here, we map for the first time genome-wide localization of endogenous mH2A1.1 and link the distribution of mH2A1.1 to control of gene expression in human breast cancer cells. In addition to localization shared with mH2A1.2 to facultative heterochromatin, mH2A1.1 specifically associates with regulatory elements required for gene activation, super-enhancers and promoters of highly expressed genes. Depending on the recruitment profile of mH2A1.1 to these elements, selective depletion of mH2A1.1 up- or downregulates its target genes. mH2A1.1 represses transcription when its binding is spread over the entire gene and promoter, and activates transcription when its binding is strictly confined to the transcription start site (TSS). Notably, RNA Polymerase II was frequently in pause at mH2A1.1-activated genes. Functionally, mH2A1.1-dependent regulation of a subset of paused genes impedes mammary tumor cell migration. Molecular mechanisms of mH2A1.1 function at the TSS uncovered by our study define an intriguing new mode of transcription regulation in cancer cells.Author SummaryControl of gene expression driving cellular functions from differentiation to epistasis and causing, when dysfunctional, uncountable diseases, relies on modifications of chromatin structure. One key element enabling chromatin plasticity is the replacement of canonical histones by histone variants. Among histone variants macroH2A1 (mH2A) is an extraordinary H2A variant possessing a large non-histone domain placed outside of the nucleosome. Two splicing isoforms, mH2A1.1 and mH2A1.2, are produced, but these are rarely studied separately because they only differ in a 30 amino acid region and are difficult to distinguish experimentally, which likely explains contradictory functions reported in the literature. Here, we take advantage of a mH2A1.1 specific antibody to generate the first genome-wide chromatin-associated map of this histone variant in the invasive breast cancer cells line MDA-MB231. We confirm that mH2A1.1, like mH2A1.2, is enriched at facultative heterochromatin in agreement with its reported role as a repressor. However, we discovered that unlike its splicing isoform, mH2A1.1 specifically binds to super-enhancers and the transcription start site of highly transcribed genes. mH2A1.1 is necessary for regulating transcription of these genes. At the cellular level, we demonstrate that mH2A1.1 inhibits migration capacity of highly metastatic breast cancer cells. Our study characterizes for the first time binding profiles of mH2A1.1 that are linked to regulation of gene expression, thereby providing a new molecular mechanisms which govern the plasticity of human tumor cells.

The Expression of Dopamine Receptors Gene and their Potential Role in Targeting Breast Cancer Cells with Selective Agonist and Antagonist Drugs. Could it be the Novel Insight to Therapy?

2019 ◽  
Vol 16 (2) ◽  
pp. 184-197 ◽  
Author(s):  
Hossein Bakhtou ◽  
Asiie Olfatbakhsh ◽  
Abdolkhaegh Deezagi ◽  
Ghasem Ahangari
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Cancer Cells ◽  
Dopamine Receptors ◽  
Breast Cancer Cells ◽  
D2 Receptors ◽  
Healthy Individuals ◽  
Agonist And Antagonist ◽  
The Mean ◽  
Mcf 7

Background:Breast cancer is one of the common causes of mortality for women in Iran and other parts of the world. The substantial increasing rate of breast cancer in both developed and developing countries warns the scientists to provide more preventive steps and therapeutic measures. This study is conducted to investigate the impact of neurotransmitters (e.g., Dopamine) through their receptors and the importance of cancers via damaging immune system. It also evaluates dopamine receptors gene expression in the women with breast cancer at stages II or III and dopamine receptor D2 (DRD2) related agonist and antagonist drug effects on human breast cancer cells, including MCF-7 and SKBR-3.Methods:The patients were categorized into two groups: 30 native patients who were diagnosed with breast cancer at stages II and III, with the mean age of 44.6 years and they were reported to have the experience of a chronic stress or unpleasant life event. The second group included 30 individuals with the mean age of 39 years as the control group. In order to determine the RNA concentration in all samples, the RNA samples were extracted and cDNA was synthesized. The MCF-7 cells and SKBR-3 cells were treated with dopamine receptors agonists and antagonists. The MTT test was conducted to identify oxidative and reductive enzymes and to specify appropriate dosage at four concentrations of dopamine and Cabergoline on MCF-7 and SKBR-3 cells. Immunofluorescence staining was done by the use of a mixed dye containing acridine orange and ethidiume bromide on account of differentiating between apoptotic and necrotic cells. Flow cytometry assay was an applied method to differentiate necrotic from apoptotic cells.Results:Sixty seven and thirty three percent of the patients were related to stages II and III, respectively. About sixty three percent of the patients expressed ER, while fifty seven percent expressed PR. Thirty seven percent of the patients were identified as HER-2 positive. All types of D2-receptors were expressed in PBMC of patients with breast cancer and healthy individuals. The expression of the whole dopamine receptor subtypes (DRD2-DRD4) was carried out on MCF-7 cell line. The results of RT-PCR confirmed the expression of DRD2 on SKBR-3 cells, whereas the other types of D2- receptors did not have an expression. The remarkable differences in gene expression rates between patients and healthy individuals were revealed in the result of the Real-time PCR analysis. The over expression in DRD2 and DRD4 genes of PBMCs was observed in the patients with breast cancer at stages II and III. The great amount of apoptosis and necrosis occurred after the treatment of MCF-7 cells by Cabergoline from 25 to 100 µmolL-1 concentrations.Conclusion:This study revealed the features of dopamine receptors associated with apoptosis induction in breast cancer cells. Moreover, the use of D2-agonist based on dopamine receptors expression in various breast tumoral cells could be promising as a new insight of complementary therapy in breast cancer.

scholarly journals Pterostilbene Changes Epigenetic Marks at Enhancer Regions of Oncogenes in Breast Cancer Cells

Antioxidants ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1232
Author(s):  
Sadaf Harandi-Zadeh ◽  
Cayla Boycott ◽  
Megan Beetch ◽  
Tony Yang ◽  
Benjamin J. E. Martin ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Cancer Cells ◽  
Breast Cancer Cells ◽  
Dna Methyltransferase ◽  
Epigenetic Marks ◽  
Chip Sequencing ◽  
Dietary Polyphenols ◽  
A Genome ◽  
Epigenetic Patterns

Epigenetic aberrations are linked to sporadic breast cancer. Interestingly, certain dietary polyphenols with anti-cancer effects, such as pterostilbene (PTS), have been shown to regulate gene expression by altering epigenetic patterns. Our group has proposed the involvement of DNA methylation and DNA methyltransferase 3B (DNMT3B) as vital players in PTS-mediated suppression of candidate oncogenes and suggested a role of enhancers as target regions. In the present study, we assess a genome-wide impact of PTS on epigenetic marks at enhancers in highly invasive MCF10CA1a breast cancer cells. Following chromatin immunoprecipitation (ChIP)-sequencing in MCF10CA1a cells treated with 7 μM PTS for 9 days, we discovered that PTS leads to increased binding of DNMT3B at enhancers of 77 genes, and 17 of those genes display an overlapping decrease in the occupancy of trimethylation at lysine 36 of histone 3 (H3K36me3), a mark of active enhancers. We selected two genes, PITPNC1 and LINC00910, and found that their enhancers are hypermethylated in response to PTS. These changes coincided with the downregulation of gene expression. Of importance, we showed that 6 out of 17 target enhancers, including PITPNC1 and LINC00910, are bound by an oncogenic transcription factor OCT1 in MCF10CA1a cells. Indeed, the six enhancers corresponded to genes with established or putative cancer-driving functions. PTS led to a decrease in OCT1 binding at those enhancers, and OCT1 depletion resulted in PITPNC1 and LINC00910 downregulation, further demonstrating a role for OCT1 in transcriptional regulation. Our findings provide novel evidence for the epigenetic regulation of enhancer regions by dietary polyphenols in breast cancer cells.

scholarly journals Estradiol Abrogates Apoptosis in Breast Cancer Cells through Inactivation of BAD: Ras-dependent Nongenomic Pathways Requiring Signaling through ERK and Akt

2004 ◽  
Vol 15 (7) ◽  
pp. 3266-3284 ◽  
Author(s):  
Romaine Ingrid Fernando ◽  
Jay Wimalasena
Keyword(s):  
Breast Cancer ◽  
Tumor Necrosis Factor ◽  
Cancer Cells ◽  
Breast Cancer Cells ◽  
Tumor Necrosis ◽  
Tumor Necrosis Factor Α ◽  
Dominant Negative ◽  
Induced Apoptosis ◽  
Factor Α ◽  
Necrosis Factor

Estrogens such as 17-β estradiol (E2) play a critical role in sporadic breast cancer progression and decrease apoptosis in breast cancer cells. Our studies using estrogen receptor-positive MCF7 cells show that E2 abrogates apoptosis possibly through phosphorylation/inactivation of the proapoptotic protein BAD, which was rapidly phosphorylated at S112 and S136. Inhibition of BAD protein expression with specific antisense oligonucleotides reduced the effectiveness of tumor necrosis factor-α, H2O2, and serum starvation in causing apoptosis. Furthermore, the ability of E2 to prevent tumor necrosis factor-α-induced apoptosis was blocked by overexpression of the BAD S112A/S136A mutant but not the wild-type BAD. BAD S112A/S136A, which lacks phosphorylation sites for p90RSK1 and Akt, was not phosphorylated in response to E2 in vitro. E2 treatment rapidly activated phosphatidylinositol 3-kinase (PI-3K)/Akt and p90RSK1 to an extent similar to insulin-like growth factor-1 treatment. In agreement with p90RSK1 activation, E2 also rapidly activated extracellular signal-regulated kinase, and this activity was down-regulated by chemical and biological inhibition of PI-3K suggestive of cross talk between signaling pathways responding to E2. Dominant negative Ras blocked E2-induced BAD phosphorylation and the Raf-activator RasV12T35S induced BAD phosphorylation as well as enhanced E2-induced phosphorylation at S112. Chemical inhibition of PI-3K and mitogen-activated protein kinase kinase 1 inhibited E2-induced BAD phosphorylation at S112 and S136 and expression of dominant negative Ras-induced apoptosis in proliferating cells. Together, these data demonstrate a new nongenomic mechanism by which E2 prevents apoptosis.

scholarly journals Curcumin inhibits leptin gene expression and secretion in breast cancer cells by estrogen receptors

2014 ◽  
Vol 14 (1) ◽  
pp. 66 ◽  
Author(s):  
Kazem Nejati-Koshki ◽  
Abolfazl Akbarzadeh ◽  
Mohammad Pourhassan-Moghaddam
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Estrogen Receptors ◽  
Cancer Cells ◽  
Breast Cancer Cells
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