LINC01977 Promotes Breast Cancer Progression and Chemoresistance to Doxorubicin by Targeting miR-212-3p/GOLM1 Axis

Long non-coding RNAs(lncRNAs) play an important role in cancer initiation and progression. However, hub lncRNAs involved in breast cancer still remain underexplored. In this study, integrated bioinformatics analysis was used to define LINC01977 as a key oncogenic driver in breast cancer. Subsequently, in vitro assays showed that LINC01977 could significantly promote breast cancer progression and chemoresistance to doxorubicin. To further investigate its biological mechanism, we performed dual-luciferase reporter assay, real-time PCR, RNA immunoprecipitation (RIP), and rescue assay. Our results indicated that LINC01977 may function as ceRNA to prevent GOLM1 gene from miRNA-mediated repression by sponging miR-212-3p. Overall, LINC01977 can serve as a novel prognostic indicator, and help develop more effective therapeutic approaches for breast cancer patients.

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miR-370-3p as a Novel Biomarker Promotes Breast Cancer Progression by Targeting FBLN5

Stem Cells International ◽

10.1155/2021/4649890 ◽

2021 ◽

Vol 2021 ◽

pp. 1-18

Author(s):

Jiahui Mao ◽

Lingxia Wang ◽

Junying Wu ◽

Yichun Wang ◽

Huiyan Wen ◽

...

Keyword(s):

Breast Cancer ◽

Cell Proliferation ◽

Cancer Patients ◽

Signaling Pathway ◽

Cancer Progression ◽

Breast Cancer Progression ◽

Breast Cancer Patients ◽

Potential Biomarker

miRNAs play a crucial part in multiple biological processes of cell proliferation, migration, apoptosis, and chemoresistance. In cancer, miRNAs can be divided into oncogenes or tumor suppressors on the basis of their functions in the carcinogenic process. The purpose of this study was to explore the roles and clinical diagnostic value of miR-370-3p in breast cancer. Our results demonstrated that miR-370-3p significantly promoted proliferation, metastasis, and stemness of breast cancer in vitro and in vivo. In particular, clinical data revealed that high expression of serum miR-370-3p and exosomal miR-370-3p from breast cancer patients was remarkably correlated with lymphatic metastasis and tumor node metastasis (TNM) stages. Mechanistically, miR-370-3p inhibited FBLN5 expression and activated the NF-κB signaling pathway to promote breast cancer cell proliferation, migration, and stemness. FBLN5 expression was significantly decreased in breast cancer cells and tumor tissues of breast cancer patients. Our research identified that miR-370-3p promoted breast cancer progression by inhibiting FBLN5 expression and activating the NF-κB signaling pathway. Serum exosomal miR-370-3p would provide a potential biomarker for the diagnosis of breast cancer.

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CircMETTL3, Upregulated in a m6A-dependent Manner, Promotes Breast Cancer Progression through circMETTL3/miR-31-5p/CDK1 axis

10.21203/rs.3.rs-125330/v1 ◽

2020 ◽

Author(s):

Zhi Li ◽

HaiYan Yang ◽

XinYuan Dai ◽

Xu Zhang ◽

YuZhou Huang ◽

...

Keyword(s):

Breast Cancer ◽

Cancer Progression ◽

Biological Function ◽

Breast Cancer Progression ◽

Host Gene ◽

Luciferase Reporter ◽

Dependent Manner ◽

Repressive Effect ◽

Rna Immunoprecipitation

Abstract Background: Growing evidence indicates N6-methyladenosine (m6A) has biological function in oncogenesis. METTL3, the catalytic component, is the most important part of methyltransferase complex and plays a crucial role in cancers. However, the biological function of circRNAs derived from METTL3 and the underlying molecular mechanism remains unclear.Methods: Quantitative real-time PCR was used to determine the circMETTL3 expression in breast cancer tissues and cell lines. Then, functional experiments in vitro and in vivo were performed to investigate the effects of circMETTL3 on tumor growth and metastasis in breast cancer. Mechanistically, fluorescent in situ hybridization, dual luciferase reporter assay, RNA pull-down and RNA immunoprecipitation experiments were performed to confirm the interaction between circMETTL3 and miR-31-5p in breast cancer. Furthermore, we explored regulatory effects of m6A modification on circMETTL3 expression with m6A RNA immunoprecipitation.Results: We found that circMETTL3 was significantly upregulated in breast cancer. The results indicated that circMETTL3 could promote breast cancer cell proliferation, migration and invasion as well as tumorigenesis in vivo. Mechanistic analysis showed that circMETTL3 might act as a ceRNA (competing endogenous RNA) of miR-31-5p to relieve the repressive effect of miR-31-5p on its target cyclin-dependent kinases (CDK1). Moreover, m6A modification of circMETTL3 might affect its expression.Conclusion: Our findings suggest that circMETTL3 promotes breast cancer progression through circMETTL3/miR-31-5p/CDK1 axis. Moreover, METTL3, the host gene of circMETTL3, may regulates circMETTL3 expression in a m6A-dependent manner, while circMETTL3 has no effect on METTL3 expression, providing a new relationship between the circRNA and the corresponding host gene. Thus, it may serve as a new diagnostic marker or therapeutic target for breast cancer patients.

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Attenuation of PITPNM1 Signaling Cascade Can Inhibit Breast Cancer Progression

Biomolecules ◽

10.3390/biom11091265 ◽

2021 ◽

Vol 11 (9) ◽

pp. 1265

Author(s):

Zihao Liu ◽

Yu Shi ◽

Qun Lin ◽

Wenqian Yang ◽

Qing Luo ◽

...

Keyword(s):

Breast Cancer ◽

Cancer Progression ◽

Molecular Taxonomy ◽

Prognostic Indicator ◽

Normal Breast ◽

The Cancer Genome Atlas ◽

Breast Cancer Patients ◽

Phosphatidylinositol Transfer ◽

Cancer Tissues

Phosphatidylinositol transfer protein membrane-associated 1 (PITPNM1) contains a highly conserved phosphatidylinositol transfer domain which is involved in phosphoinositide trafficking and signaling transduction under physiological conditions. However, the functional role of PITPNM1 in cancer progression remains unknown. Here, by integrating datasets of The Cancer Genome Atlas (TCGA) and Molecular Taxonomy of Breast Cancer (METABRIC), we found that the expression of PITPNM1 is much higher in breast cancer tissues than in normal breast tissues, and a high expression of PITPNM1 predicts a poor prognosis for breast cancer patients. Through gene set variation analysis (GSEA) and gene ontology (GO) analysis, we found PITPNM1 is mainly associated with carcinogenesis and cell-to-cell signaling ontology. Silencing of PITPNM1, in vitro, significantly abrogates proliferation and colony formation of breast cancer cells. Collectively, PITPNM1 is an important prognostic indicator and a potential therapeutic target for breast cancer.

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Significance of FGFR2 – RSK2 interdependence for breast cancer progression. In vitro studies and clinical material analyses

Postępy Polskiej Medycyny i Farmacji ◽

10.5604/01.3001.0013.2321 ◽

2019 ◽

Vol 6 ◽

pp. 77-83

Author(s):

Dominika Czaplińska

Keyword(s):

Breast Cancer ◽

Cancer Progression ◽

Focal Adhesions ◽

Disease Free Survival ◽

Breast Cancer Progression ◽

Signalling Pathway ◽

In Vitro Studies ◽

Breast Cancer Patients ◽

Ribosomal S6 Kinase

The members of p90 ribosomal S6 kinase (RSK) family of Ser/Thr kinases are downstream effectors of MAPK/ERK pathway that regulate diverse cellular processes including cell growth, proliferation and survival. In carcinogenesis, RSKs are thought to modulate cell motility, invasion and metastasis. Elevated level of RSK1 and RSK2 was found in about 50% of breast tumours and is associated with a poor survival outcome. The project aims to investigate the role of RSK2 kinase in breast cancer progression out at two complementary levels: in vitro studies and analyses of samples from breast cancer patients. We demonstrated that in breast epithelial and cancer cell lines stimulation of FGFR2 (fibroblast growth factor receptor 2) with FGF2 induced activation of RSK2-Tyr529 and that this new signalling pathway promoted anchorage-independent cell proliferation, formation of focal adhesions and cell migration. In clinical analyses FGFR2 showed a positive correlation with RSK2 at both protein (p = 0.003) and mRNA (p = 0.001) levels. The values of correlation coefficients were the highest in triple- -negative breast cancer (TNBC), which is more aggressive and has a poorer prognosis than other breast cancer subtypes. Moreover, expression of FGFR2 and/or activated RSK (RSK-P) significantly correlated with poor disease-free survival (DFS) of patients (p = 0.01). Taken together, our results suggest that FGFR2-RSK2 signalling pathway is associated with breast cancer progression and further studies of FGFR2-RSK2 interdependence may contribute to the development of new therapeutic strategies in the management of patients with breast cancer.

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Twist-mediated PAR1 induction is required for breast cancer progression and metastasis by inhibiting Hippo pathway

Cell Death and Disease ◽

10.1038/s41419-020-2725-4 ◽

2020 ◽

Vol 11 (7) ◽

Author(s):

Yifan Wang ◽

Ruocen Liao ◽

Xingyu Chen ◽

Xuhua Ying ◽

Guanping Chen ◽

...

Keyword(s):

Breast Cancer ◽

Cancer Cells ◽

Cancer Progression ◽

Invasive Breast Cancer ◽

Breast Cancer Cells ◽

Epithelial Mesenchymal Transition ◽

Hippo Pathway ◽

Breast Cancer Patients ◽

Mesenchymal Transition

Abstract Breast cancer is considered to be the most prevalent cancer in women worldwide, and metastasis is the primary cause of death. Protease-activated receptor 1 (PAR1) is a GPCR family member involved in the invasive and metastatic processes of cancer cells. However, the functions and underlying mechanisms of PAR1 in breast cancer remain unclear. In this study, we found that PAR1 is highly expressed in high invasive breast cancer cells, and predicts poor prognosis in ER-negative and high-grade breast cancer patients. Mechanistically, Twist transcriptionally induces PAR1 expression, leading to inhibition of Hippo pathway and activation of YAP/TAZ; Inhibition of PAR1 suppresses YAP/TAZ-induced epithelial-mesenchymal transition (EMT), invasion, migration, cancer stem cell (CSC)-like properties, tumor growth and metastasis of breast cancer cells in vitro and in vivo. These findings suggest that PAR1 acts as a direct transcriptionally target of Twist, can promote EMT, tumorigenicity and metastasis by controlling the Hippo pathway; this may lead to a potential therapeutic target for treating invasive breast cancer.

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Enhancers mapping uncovers phenotypic heterogeneity and evolution in patients with luminal breast cancer

10.1101/193771 ◽

2017 ◽

Author(s):

Darren K. Patten ◽

Giacomo Corleone ◽

Balázs Győrffy ◽

Edina Erdős ◽

Alina Saiakhova ◽

...

Keyword(s):

Breast Cancer ◽

Cancer Progression ◽

Regulatory Elements ◽

Phenotypic Heterogeneity ◽

Endocrine Treatment ◽

Clinical Samples ◽

Luminal Breast Cancer ◽

Breast Cancer Patients ◽

Expression Of Genes

AbstractThe degree of intrinsic and interpatient phenotypic heterogeneity and its role in tumour evolution is poorly understood. Phenotypic divergence can be achieved via the inheritance of alternative transcriptional programs1,2. Cell-type specific transcription is maintained through the activation of epigenetically-defined regulatory regions including promoters and enhancers1,3,4. In this work, we annotated the epigenome of 47 primary and metastatic oestrogen-receptor (ERα)-positive breast cancer specimens from clinical samples, and developed strategies to deduce phenotypic heterogeneity from the regulatory landscape, identifying key regulatory elements commonly shared across patients. Highly shared regions contain a unique set of regulatory information including the motif for the transcription factor YY1. In vitro work shows that YY1 is essential for ERα transcriptional activity and defines the critical subset of functional ERα binding sites driving tumor growth in most luminal patients. YY1 also control the expression of genes that mediate resistance to endocrine treatment. Finally, we show that H3K27ac levels at active enhancer elements can be used as a surrogate of intra-tumor phenotypic heterogeneity, and to track expansion and contraction of phenotypic subpopulations throughout breast cancer progression. Tracking YY1 and SLC9A3R1 positive clones in primary and metastatic lesions, we show that endocrine therapies drive the expansion of phenotypic clones originally underrepresented at diagnosis. Collectively, our data show that epigenetic mechanisms significantly contribute to phenotypic heterogeneity and evolution in systemically treated breast cancer patients.

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CircBCBM1 Promotes Breast Cancer Brain Metastasis via miR-125a/BRD4 Axis

10.21203/rs.3.rs-130879/v1 ◽

2020 ◽

Author(s):

Bo Fu ◽

Wei Liu ◽

Peng Li ◽

Li Pan ◽

Ke Li ◽

...

Keyword(s):

Breast Cancer ◽

Brain Metastasis ◽

Luciferase Reporter ◽

Circular Rnas ◽

Breast Cancer Patients ◽

Breast Cancer Brain Metastasis ◽

Tumorigenesis And Progression ◽

And Migration

Abstract Background: Accumulating evidence indicates that circular RNAs (circRNAs) play critical roles in tumorigenesis and progression of various cancers. We previously identified a novel upregulated circRNA, circBCBM1 (hsa_circ_0001944), in the context of breast cancer brain metastasis. However, the potential biological function and molecular mechanism of circBCBM1 in breast cancer brain metastasis remain largely unknown.Methods: In this reserch, we validated the expression and characterization of circBCBM1 through RT-qPCR, Sanger sequencing, RNase R assay and fluorescence in situ hybridization (FISH). Functional experiments were performed to determine the effect of circBCBM1 on growth and metastasis of 231-BR cells both in vitro and in vivo. The regulatory mechanisms among circBCBM1, miR-125a (has-miR-125a-5p), and BRD4 (bromodomain containing 4) were investigated by RNA immunoprecipitation (RIP), RNA pull-down, luciferase reporter assay and western blot. Results: Our findings demonstrated that circBCBM1 is a stable and cytoplasmic circRNA. Functionally, silencing of circBCBM1 led to decreased proliferation and migration of 231-BR cells whereas elevated circBCBM1 expression showed reverse effects in vitro. These findings were confirmed in vivo in mouse models, as knockdown of circBCBM1 significantly decreased growth and brain metastases of 231-BR cells. Mechanistically, circBCBM1 functions as an endogenous miR-125a sponge to inhibit miR-125a activity, resulting in the upregulation of BRD4 expression and subsequent upregulation of MMP9 (matrix metallopeptidase 9) through Sonic hedgehog (SHH) signaling pathway. Importantly, circBCBM1 was markedly upregulated in the breast cancer brain metastasis cells and clinical tissue and plasma samples; besides, the overexpression of circBCBM1 in primary cancerous tissues was associated with shorter brain metastasis-free survival (BMFS) of breast cancer patients.Conclusions: These findings indicate that circBCBM1 is involved in breast cancer brain metastasis via circBCBM1/miR-125a/BRD4 axis, which sheds light on the pathogenic mechanism of circBCBM1 and provides translational evidence that circBCBM1 may serve as a novel diagnostic or prognostic biomarker and potential therapeutic target for breast cancer brain metastasis.

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The GATA3 X308_Splice breast cancer mutation is a hormone context-dependent oncogenic driver

Oncogene ◽

10.1038/s41388-020-1376-3 ◽

2020 ◽

Vol 39 (32) ◽

pp. 5455-5467

Author(s):

Natascha Hruschka ◽

Mark Kalisz ◽

Maria Subijana ◽

Osvaldo Graña-Castro ◽

Francisco Del Cano-Ochoa ◽

...

Keyword(s):

Breast Cancer ◽

Progesterone Receptors ◽

Transcriptional Response ◽

Molecular Data ◽

Fine Tuning ◽

Driver Mutations ◽

Breast Cancer Patients ◽

Oncogenic Driver ◽

Functional Analyses

Abstract As the catalog of oncogenic driver mutations is expanding, it becomes clear that alterations in a given gene might have different functions and should not be lumped into one class. The transcription factor GATA3 is a paradigm of this. We investigated the functions of the most common GATA3 mutation (X308_Splice) and five additional mutations, which converge into a neoprotein that we called “neoGATA3,” associated with excellent prognosis in patients. Analysis of available molecular data from >3000 breast cancer patients revealed a dysregulation of the ER-dependent transcriptional response in tumors carrying neoGATA3-generating mutations. Mechanistic studies in vitro showed that neoGATA3 interferes with the transcriptional programs controlled by estrogen and progesterone receptors, without fully abrogating them. ChIP-Seq analysis indicated that ER binding is reduced in neoGATA3-expressing cells, especially at distal regions, suggesting that neoGATA3 interferes with the fine tuning of ER-dependent gene expression. This has opposite outputs in distinct hormonal context, having pro- or anti-proliferative effects, depending on the estrogen/progesterone ratio. Our data call for functional analyses of putative cancer drivers to guide clinical application.

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LncRNA NEAT1 accelerates breast cancer progression through regulating miR-410-3p/ CCND1 axis

Cancer Biomarkers ◽

10.3233/cbm-190721 ◽

2020 ◽

Vol 29 (2) ◽

pp. 277-290

Author(s):

Xuan Liu ◽

Weirong Yao ◽

Haiwei Xiong ◽

Qiang Li ◽

Yingliang Li

Keyword(s):

Breast Cancer ◽

Tumor Growth ◽

Cancer Cells ◽

Cancer Progression ◽

Breast Cancer Cells ◽

Breast Cancer Progression ◽

Cell Counting ◽

Luciferase Reporter ◽

Cancer Tissues

BACKGROUND: Breast cancer is the most common malignant tumor and usually occurs in women. Studies have shown that lncRNA nuclear enriched abundant transcript 1 (NEAT1) contributes to breast cancer progression. This study intends to further investigate the molecular mechanism of NEAT1 in breast cancer. METHODS: The expression levels of NEAT1, miR-410-3p and Cyclin D1 (CCND1) were detected by quantitative real-time PCR (qRT-PCR) in breast cancer tissues and cells. Kaplan-Meier analysis and the log-rank test were performed to determine the relationship between NEAT1 and overall survival. Cell Counting Kit-8 (CCK-8) assay analyzed cell proliferation. Transwell assay was performed to examine cell migration and invasion. The protein levels of CCND1 and epithelial-mesenchymal transition (EMT)-related proteins (E-cadherin, N-cadherin and Vimentin) were measured by western blot. The target relationship was predicted by bioinformatics analysis, and confirmed by luciferase reporter assay and RNA Immunoprecipitation (RIP) assay. Xenograft analysis was used to evaluate the tumor growth in vivo. RESULTS: NEAT1 and CCND1 were upregulated, while miR-410-3p was down-regulated in breast cancer tissues and cells. Higher NEAT1 expression level was associated with lower survival rate of breast cancer patients. Knockdown of miR-410-3p restored silenced NEAT1-mediated the inhibition of on proliferation, migration, invasion and EMT of breast cancer cells. In addition, NEAT1 regulated CCND1 expression by sponging miR-410-3p in breast cancer cells. NEAT1 knockdown blocked the tumor growth in vivo. CONCLUSION: NEAT1 induced breast cancer progression by regulating the miR-410-3p/CCND1 axis, indicating that NEAT1 may be a potential therapeutic target in breast cancer.

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ASPH-notch Axis guided Exosomal delivery of Prometastatic Secretome renders breast Cancer multi-organ metastasis

Molecular Cancer ◽

10.1186/s12943-019-1077-0 ◽

2019 ◽

Vol 18 (1) ◽

Cited By ~ 8

Author(s):

Qiushi Lin ◽

Xuesong Chen ◽

Fanzheng Meng ◽

Kosuke Ogawa ◽

Min Li ◽

...

Keyword(s):

Breast Cancer ◽

Molecular Mechanisms ◽

Ductal Carcinoma ◽

Disease Free Survival ◽

Normal Adult ◽

Luciferase Reporter ◽

Breast Cancer Patients ◽

Long Distance ◽

Premetastatic Niche

Abstract Background Aspartate β-hydroxylase (ASPH) is silent in normal adult tissues only to re-emerge during oncogenesis where its function is required for generation and maintenance of malignant phenotypes. Exosomes enable prooncogenic secretome delivering and trafficking for long-distance cell-to-cell communication. This study aims to explore molecular mechanisms underlying how ASPH network regulates designated exosomes to program development and progression of breast cancer. Methods Stable cell lines overexpressing or knocking-out of ASPH were established using lentivirus transfection or CRISPR-CAS9 systems. Western blot, MTT, immunofluorescence, luciferase reporter, co-immunoprecipitation, 2D/3-D invasion, tube formation, mammosphere formation, immunohistochemistry and newly developed in vitro metastasis were applied. Results Through physical interactions with Notch receptors, ligands (JAGs) and regulators (ADAM10/17), ASPH activates Notch cascade to provide raw materials (especially MMPs/ADAMs) for synthesis/release of pro-metastatic exosomes. Exosomes orchestrate EMT, 2-D/3-D invasion, stemness, angiogenesis, and premetastatic niche formation. Small molecule inhibitors (SMIs) of ASPH’s β-hydroxylase specifically/efficiently abrogated in vitro metastasis, which mimics basement membrane invasion at primary site, intravasation/extravasation (transendothelial migration), and colonization/outgrowth at distant sites. Multiple organ-metastases in orthotopic and tail vein injection murine models were substantially blocked by a specific SMI. ASPH is silenced in normal adult breast, upregulated from in situ malignancies to highly expressed in invasive/advanced ductal carcinoma. Moderate-high expression of ASPH confers more aggressive molecular subtypes (TNBC or Her2 amplified), early recurrence/progression and devastating outcome (reduced overall/disease-free survival) of breast cancer. Expression profiling of Notch signaling components positively correlates with ASPH expression in breast cancer patients, confirming that ASPH-Notch axis acts functionally in breast tumorigenesis. Conclusions ASPH-Notch axis guides particularly selective exosomes to potentiate multifaceted metastasis. ASPH’s pro-oncogenic/pro-metastatic properties are essential for breast cancer development/progression, revealing a potential target for therapy.

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