SETD5 Binding to EP300/HIF1α is Required for Glycolysis in Breast Cancer Stem-Like Cells

Abstract BackgroundGlycolysis plays a pivotal role in breast cancer stem-like cell reprogramming. The SET-domain containing 5 (SETD5) is a previously uncharacterized member of the histone lysine methyltransferase family. Yet, the molecular mechanisms underlying the promotion of stem-like and glycolysis activation traits of SETD5 have not been elucidated.MethodsBasing on public datasets, we explored clinicopathological and survival analysis of SETD5 on breast cancer (BC) patients. Spheroid formation, transfection experiments and measurement of glucose uptake and lactate production analyzed the regulatory function of SETD5 on glycolysis in breast cancer stem-like cells (BCSC). The impact of SETD5 on tumor growth was studied in a murine xenograft model. Immunohistochemistry, immunofluorescence, western blot, preparation of cytoplasmic and nuclear extracts and co-immunoprecipitation were used to determine the molecular mechanisms of SETD5 in cancer cell glycolysis.ResultsOur data displayed that overexpression of SETD5 in BC tissues is positively associated with progression. SETD5 overexpression is associated with poor post-progression survival in BC patients. SETD5 expression was enriched in spheroid cells. Downregulation of SETD5 significantly decreased BCSC properties and glycolysis in vitro and in vivo. Interestingly, SETD5 and glycolytic enzymes were accumulated in the central hypoxic regions of subcutaneous tumor tissues. Our mechanistic study found that SETD5 binding to EP300/hypoxia-inducible factor 1α (HIF1α) and work as an upstream effector. SETD5 knockdown reduced the expression of HIF1α, hexokinase-2, and 6-phosphofructo-2-kinase in the nucleus after treatment with cobalt chloride (CoCl2), a chemical hypoxia mimetic agent, which activates HIF1α to accumulate in the nucleus. ConclusionSETD5 is required for glycolysis in BCSCs through binding to EP300/HIF1α and could be a potential therapeutic target for BC patients.

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CLDN6 Suppresses c–MYC–Mediated Aerobic Glycolysis to Inhibit Proliferation by TAZ in Breast Cancer

International Journal of Molecular Sciences ◽

10.3390/ijms23010129 ◽

2021 ◽

Vol 23 (1) ◽

pp. 129

Author(s):

Huinan Qu ◽

Da Qi ◽

Xinqi Wang ◽

Yuan Dong ◽

Qiu Jin ◽

...

Keyword(s):

Breast Cancer ◽

Molecular Mechanisms ◽

Aerobic Glycolysis ◽

Lactate Production ◽

Suppressor Gene ◽

Binding Motif ◽

Cancer Proliferation ◽

In Vivo Experiments

Claudin 6 (CLDN6) was found to be a breast cancer suppressor gene, which is lowly expressed in breast cancer and inhibits breast cancer cell proliferation upon overexpression. However, the mechanism by which CLDN6 inhibits breast cancer proliferation is unclear. Here, we investigated this issue and elucidated the molecular mechanisms by which CLDN6 inhibits breast cancer proliferation. First, we verified that CLDN6 was lowly expressed in breast cancer tissues and that patients with lower CLDN6 expression had a worse prognosis. Next, we confirmed that CLDN6 inhibited breast cancer proliferation through in vitro and in vivo experiments. As for the mechanism, we found that CLDN6 inhibited c–MYC–mediated aerobic glycolysis based on a metabolomic analysis of CLDN6 affecting cellular lactate levels. CLDN6 interacted with a transcriptional co–activator with PDZ-binding motif (TAZ) and reduced the level of TAZ, thereby suppressing c–MYC transcription, which led to a reduction in glucose uptake and lactate production. Considered together, our results suggested that CLDN6 suppressed c–MYC–mediated aerobic glycolysis to inhibit the proliferation of breast cancer by TAZ, which indicated that CLDN6 acted as a novel regulator of aerobic glycolysis and provided a theoretical basis for CLDN6 as a biomarker of progression in breast cancer.

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SMC1A promotes tumor growth in breast cancer by activation of the AKT/FOXM1/STMN1 Signaling Pathway

10.21203/rs.3.rs-861071/v1 ◽

2021 ◽

Author(s):

kaichun li ◽

Ping Dai ◽

Lin Xue ◽

Long Liu ◽

Shiyu Cheng ◽

...

Keyword(s):

Breast Cancer ◽

Molecular Mechanisms ◽

Xenograft Model ◽

Cell Cycle Checkpoints ◽

Biological Functions ◽

Tumor Tissues ◽

Subcutaneous Xenograft ◽

Novel Target

Abstract Background SMC1A (Structural maintenance of chromosomes 1) is overexpressed in various cancers and acts as an oncogene which has been implicated in critical biological functions (cell-cycle checkpoints regulation, cell division, and DNA repair). However, the mechanism and role of SMC1A in breast cancer are poorly understood. Methods TCGA database was utilized to explore the expression of SMC1A and the relationship between SMC1A and FOXM1 and STMN1. Subsequently, short hairpin RNA (shRNA) targeting SMC1A was used to examined the biological functions of it in MDA-MB-231 and MDA-MB-468 cells. Finally, subcutaneous xenograft model to verify the roles of SMC1A in vivo. Results In the present study, we demonstrated that SMC1A was significantly increased in breast cancer (BC) via TCGA database. Then loss and gain of function studies revealed that SMC1A contributed to BC cell survival, apoptosis, and invasion. Interestingly, we found that SMC1A triggered the AKT/FOXM1 cascade, which promoted BC cell proliferation. Furthermore, overexpression of FOXM1 abolished the inhibition of cell growth induced by SMC1A silencing in vitro. Clinically, the expression of SMC1A in BC tumor tissues is positively correlated with the expression of FOXM1. Conclusion Taken together, our findings not only enhanced our understanding of molecular mechanisms of SMC1A in BC, but also might provide a novel target for the development of therapeutic strategies.

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Decitabine Reverses CSC-Induced Docetaxel Resistance via Epigenetic Regulation of DAB2IP in TNBC.

10.21203/rs.3.rs-51242/v1 ◽

2020 ◽

Author(s):

Zhenchong Xiong ◽

Lin Yang ◽

Lu Yang ◽

Jianchang Fu ◽

Peng Liu ◽

...

Keyword(s):

Breast Cancer ◽

Cell Lines ◽

Molecular Mechanisms ◽

Low Dose ◽

Methylation Status ◽

Pcr Analysis ◽

Tnbc Cell ◽

The Impact

Abstract Background: Although docetaxel (DOC)-based chemotherapy, the standard of care for triple-negative breast cancer (TNBC), has greatly improved cancer survival, chemoresistance invariably evolves. Methods: Using TNBC and docetaxel-resistant TNBC cell lines, we established murine breast cancer xenograft models to evaluate the impact of decitabine on DOC resistance. The methylation status of DAB2IP was evaluated in 226 TNBC patients and TNBC cell lines by BSP analysis. A limiting dilution assay was performed to evaluate the tumorigenic capacity in vivo. Cellular and molecular mechanisms were demonstrated using in vivo and in vitro biochemical methods.Results: Here, we observed that a low dose of DAC significantly prevented the evolution of DOC resistance via inhibition of cancer stem cell (CSC) enrichment in TNBC. The bisulfite sequencing PCR analysis showed that DAC treatment increased DAB2IP expression via inhibition of DNA methylation in both DOC-resistant TNBC cells and tumorspheres. Loss of DAB2IP boosted the enrichment of CSCs through activation of Erk/β-catenin signaling in vitro and facilitated tumor initiation in vivo. In mouse model, TNBC tumors with inhibition of DAB2IP exhibited poor response to DOC and DOC-resistant tumors were resensitized to DOC treatment by low-dose DAC treatment in vivo. Finally, hypermethylation of DAB2IP was correlated with low expression of DAB2IP and poor outcome of TNBC patients treated with DOC-based chemotherapy. Conclusions: Collectively, we discovered that epigenetic silencing of DAB2IP is a driver of CSC enrichment, which results in the evolution of DOC resistance, and a low dose of DAC reverses DOC resistance through epigenetic re-expression of DAB2IP.

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The impact of FGF19/FGFR4 signaling inhibition in antitumor activity of multi-kinase inhibitors in hepatocellular carcinoma

Scientific Reports ◽

10.1038/s41598-021-84117-9 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Hiroaki Kanzaki ◽

Tetsuhiro Chiba ◽

Junjie Ao ◽

Keisuke Koroki ◽

Kengo Kanayama ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Molecular Mechanisms ◽

Kinase Inhibitors ◽

Progression Free Survival ◽

Erk Signaling ◽

Enzyme Linked Immunosorbent Assay ◽

Antitumor Effects ◽

The Impact

AbstractFGF19/FGFR4 autocrine signaling is one of the main targets for multi-kinase inhibitors (MKIs). However, the molecular mechanisms underlying FGF19/FGFR4 signaling in the antitumor effects to MKIs in hepatocellular carcinoma (HCC) remain unclear. In this study, the impact of FGFR4/ERK signaling inhibition on HCC following MKI treatment was analyzed in vitro and in vivo assays. Serum FGF19 in HCC patients treated using MKIs, such as sorafenib (n = 173) and lenvatinib (n = 40), was measured by enzyme-linked immunosorbent assay. Lenvatinib strongly inhibited the phosphorylation of FRS2 and ERK, the downstream signaling molecules of FGFR4, compared with sorafenib and regorafenib. Additional use of a selective FGFR4 inhibitor with sorafenib further suppressed FGFR4/ERK signaling and synergistically inhibited HCC cell growth in culture and xenograft subcutaneous tumors. Although serum FGF19high (n = 68) patients treated using sorafenib exhibited a significantly shorter progression-free survival and overall survival than FGF19low (n = 105) patients, there were no significant differences between FGF19high (n = 21) and FGF19low (n = 19) patients treated using lenvatinib. In conclusion, robust inhibition of FGF19/FGFR4 is of importance for the exertion of antitumor effects of MKIs. Serum FGF19 levels may function as a predictive marker for drug response and survival in HCC patients treated using sorafenib.

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Edelfosine nanoemulsions inhibit tumor growth of triple negative breast cancer in zebrafish xenograft model

Scientific Reports ◽

10.1038/s41598-021-87968-4 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Sofia M. Saraiva ◽

Carlha Gutiérrez-Lovera ◽

Jeannette Martínez-Val ◽

Sainza Lores ◽

Belén L. Bouzo ◽

...

Keyword(s):

Breast Cancer ◽

Tumor Growth ◽

Triple Negative Breast Cancer ◽

Triple Negative ◽

Xenograft Model ◽

Skin Barrier ◽

Zebrafish Embryos ◽

Biorelevant Media

AbstractTriple negative breast cancer (TNBC) is known for being very aggressive, heterogeneous and highly metastatic. The standard of care treatment is still chemotherapy, with adjacent toxicity and low efficacy, highlighting the need for alternative and more effective therapeutic strategies. Edelfosine, an alkyl-lysophospholipid, has proved to be a promising therapy for several cancer types, upon delivery in lipid nanoparticles. Therefore, the objective of this work was to explore the potential of edelfosine for the treatment of TNBC. Edelfosine nanoemulsions (ET-NEs) composed by edelfosine, Miglyol 812 and phosphatidylcholine as excipients, due to their good safety profile, presented an average size of about 120 nm and a neutral zeta potential, and were stable in biorelevant media. The ability of ET-NEs to interrupt tumor growth in TNBC was demonstrated both in vitro, using a highly aggressive and invasive TNBC cell line, and in vivo, using zebrafish embryos. Importantly, ET-NEs were able to penetrate through the skin barrier of MDA-MB 231 xenografted zebrafish embryos, into the yolk sac, leading to an effective decrease of highly aggressive and invasive tumoral cells’ proliferation. Altogether the results demonstrate the potential of ET-NEs for the development of new therapeutic approaches for TNBC.

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High expression of PSMC2 promotes gallbladder cancer through regulation of GNG4 and predicts poor prognosis

Oncogenesis ◽

10.1038/s41389-021-00330-1 ◽

2021 ◽

Vol 10 (5) ◽

Author(s):

Dawei Zhu ◽

Xing Gu ◽

Zhengyu Lin ◽

Dandan Yu ◽

Jing Wang

Keyword(s):

Gallbladder Cancer ◽

Malignant Tumor ◽

Xenograft Model ◽

Tumor Grade ◽

Significant Inhibition ◽

Mechanistic Study ◽

Downstream Target ◽

Advanced Tumor

AbstractGallbladder cancer (GBC) is a common malignant tumor of the biliary tract, which accounts for 80–95% of biliary tumors worldwide, and is the leading cause of biliary malignant tumor-related death. This study identified PSMC2 as a potential regulator in the development of GBC. We showed that PSMC2 expression in GBC tissues is significantly higher than that in normal tissues, while high PSMC2 expression was correlated with more advanced tumor grade and poorer prognosis. The knockdown of PSMC2 in GBC cells induced significant inhibition of cell proliferation, colony formation and cell motility, while the promotion of cell apoptosis. The construction and observation of the mice xenograft model also confirmed the inhibitory effects of PSMC2 knockdown on GBC development. Moreover, our mechanistic study recognized GNG4 as a potential downstream target of PSMC2, knockdown of which could aggravate the tumor suppression induced by PSMC2 knockdown in vitro and in vivo. In conclusion, for the first time, PSMC2 was revealed as a tumor promotor in the development of GBC, which could regulate cell phenotypes of GBC cells through the interaction with GNG4, and maybe a promising therapeutic target in GBC treatment.

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Effect of Auraptene on angiogenesis in Xenograft model of breast cancer

Hormone Molecular Biology and Clinical Investigation ◽

10.1515/hmbci-2021-0056 ◽

2021 ◽

Vol 0 (0) ◽

Author(s):

Mohammad Reza Shiran ◽

Elham Mahmoudian ◽

Abolghasem Ajami ◽

Seyed Mostafa Hosseini ◽

Ayjamal Khojasteh ◽

...

Keyword(s):

Breast Cancer ◽

Gene Expression ◽

Animal Model ◽

Protein Expression ◽

Western Blot ◽

Xenograft Model ◽

Dependent Manner ◽

Concentration Dependent Manner

Abstract Objectives Angiogenesis is the most important challenge in breast cancer treatment. Recently, scientists become interesting in rare natural products and intensive researches was performed to identify their pharmacological profile. Auraptene shows helpful effects such as cancer chemo-preventive, anti-inflammatory, anti-oxidant, immuno-modulatory. In this regard, we investigated the anti-angiogenesis effect of Auraptene in in-vitro and in-vivo model of breast cancer. Methods In this study, 4T, MDA-MB-231 and HUVEC cell lines were used. The proliferation study was done by MTT assay. For tube formation assay, 250 matrigel, 1 × 104 HUVEC treated with Auraptene, 20 ng/mL EGF, 20 ng/mL bFGF and 20 ng/mL VEGF were used. Gene expression of important gene related to angiogenesis in animal model of breast cancer was investigated by Real-time PCR. Protein expression of VCAM-1 and TNFR-1 gene related to angiogenesis in animal model of breast cancer was investigated by western-blot. Results Auraptene treatment led to reduction in cell viability of MDA-MB-231 in a concentration-dependent manner. Also, we observed change in the number of tubes or branches formed by cells incubated with 40 and 80 μM Auraptene. Auraptene effect the gene expression of important gene related to angiogenesis (VEGF, VEGFR2, COX2, IFNɣ). Moreover, the western blot data exhibited that Auraptene effect the protein expression of VCAM-1 and TNFR-1. Conclusions Overall, this study shows that Auraptene significantly suppressed angiogenesis via down-regulation of VEGF, VEGFR2, VCAM-1, TNFR-1, COX-2 and up-regulation of IFNγ.

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Isoform-specific promotion of breast cancer tumorigenicity by TBX3 involves induction of angiogenesis

Laboratory Investigation ◽

10.1038/s41374-019-0326-6 ◽

2019 ◽

Vol 100 (3) ◽

pp. 400-413

Author(s):

Milica Krstic ◽

Haider M. Hassan ◽

Bart Kolendowski ◽

M. Nicole Hague ◽

Pieter. H. Anborgh ◽

...

Keyword(s):

Breast Cancer ◽

Molecular Subtype ◽

Xenograft Model ◽

Tumor Formation ◽

Transcriptional Analysis ◽

Breast Cancer Cell Lines ◽

Tumor Vascularity ◽

Mouse Xenograft Model

Abstract TBX3 is a member of the highly conserved family of T-box transcription factors involved in embryogenesis, organogenesis and tumor progression. While the functional role of TBX3 in tumorigenesis has been widely studied, less is known about the specific functions of the different isoforms (TBX3iso1 and TBX3iso2) which differ in their DNA-binding domain. We therefore sought to investigate the functional consequence of this highly conserved splice event as it relates to TBX3-induced tumorigenesis. By utilizing a nude mouse xenograft model, we have identified differential tumorigenic potential between TBX3 isoforms, with TBX3iso1 overexpression more commonly associated with invasive carcinoma and high tumor vascularity. Transcriptional analysis of signaling pathways altered by TBX3iso1 and TBX3iso2 overexpression revealed significant differences in angiogenesis-related genes. Importantly, osteopontin (OPN), a cancer-associated secreted phosphoprotein, was significantly up-regulated with TBX3iso1 (but not TBX3iso2) overexpression. This pattern was observed across three non/weakly-tumorigenic breast cancer cell lines (21PT, 21NT, and MCF7). Up-regulation of OPN in TBX3iso1 overexpressing cells was associated with induction of hyaluronan synthase 2 (HAS2) expression and increased retention of hyaluronan in pericellular matrices. These transcriptional changes were accompanied by the ability to induce endothelial cell vascular channel formation by conditioned media in vitro, which could be inhibited through addition of an OPN neutralizing antibody. Within the TCGA breast cancer cohort, we identified an 8.1-fold higher TBX3iso1 to TBX3iso2 transcript ratio in tumors relative to control, and this ratio was positively associated with high-tumor grade and an aggressive molecular subtype. Collectively, the described changes involving TBX3iso1-dependent promotion of angiogenesis may thus serve as an adaptive mechanism within breast cancer cells, potentially explaining differences in tumor formation rates between TBX3 isoforms in vivo. This study is the first of its kind to report significant functional differences between the two TBX3 isoforms, both in vitro and in vivo.

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Knockdown of Leptin Receptor Affects Macrophage Phenotype in the Tumor Microenvironment Inhibiting Breast Cancer Growth and Progression

Cancers ◽

10.3390/cancers12082078 ◽

2020 ◽

Vol 12 (8) ◽

pp. 2078

Author(s):

Luca Gelsomino ◽

Giuseppina Daniela Naimo ◽

Rocco Malivindi ◽

Giuseppina Augimeri ◽

Salvatore Panza ◽

...

Keyword(s):

Breast Cancer ◽

Tumor Microenvironment ◽

Leptin Receptor ◽

Macrophage Phenotype ◽

In Vivo Models ◽

Monocyte Chemoattractant Protein 1 ◽

Arginase 1 ◽

The Impact

Aberrant leptin (Ob) signaling, a hallmark of obesity, has been recognized to influence breast cancer (BC) biology within the tumor microenvironment (TME). Here, we evaluated the impact of leptin receptor (ObR) knockdown in affecting BC phenotype and in mediating the interaction between tumor cells and macrophages, the most abundant immune cells within the TME. The stable knockdown of ObR (ObR sh) in ERα-positive and ERα-negative BC cells turned the tumor phenotype into a less aggressive one, as evidenced by in vitro and in vivo models. In xenograft tumors and in co-culture experiments between circulating monocytes and BC cells, the absence of ObR reduced the recruitment of macrophages, and also affected their cytokine mRNA expression profile. This was associated with a decreased expression and secretion of monocyte chemoattractant protein-1 in ObR sh clones. The loss of Ob/ObR signaling modulated the immunosuppressive TME, as shown by a reduced expression of programmed death ligand 1/programmed cell death protein 1/arginase 1. In addition, we observed increased phagocytic activity of macrophages compared to control Sh clones in the presence of ObR sh-derived conditioned medium. Our findings, addressing an innovative role of ObR in modulating immune TME, may open new avenues to improve BC patient health care.

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Can Hypoxic Conditioning Improve Bone Metabolism? A Systematic Review

International Journal of Environmental Research and Public Health ◽

10.3390/ijerph16101799 ◽

2019 ◽

Vol 16 (10) ◽

pp. 1799 ◽

Cited By ~ 7

Author(s):

Marta Camacho-Cardenosa ◽

Alba Camacho-Cardenosa ◽

Rafael Timón ◽

Guillermo Olcina ◽

Pablo Tomas-Carus ◽

...

Keyword(s):

Bone Metabolism ◽

Critical Role ◽

Hypoxia Inducible Factor ◽

Experimental Studies ◽

Hormonal Responses ◽

Hypoxia Exposure ◽

Ambient Oxygen ◽

The Impact

Among other functions, hypoxia-inducible factor plays a critical role in bone–vascular coupling and bone formation. Studies have suggested that hypoxic conditioning could be a potential nonpharmacological strategy for treating skeletal diseases. However, there is no clear consensus regarding the bone metabolism response to hypoxia. Therefore, this review aims to examine the impact of different modes of hypoxia conditioning on bone metabolism. The PubMed and Web of Science databases were searched for experimental studies written in English that investigated the effects of modification of ambient oxygen on bone remodelling parameters of healthy organisms. Thirty-nine studies analysed the effect of sustained or cyclic hypoxia exposure on genetic and protein expression and mineralisation capacity of different cell models; three studies carried out in animal models implemented sustained or cyclic hypoxia; ten studies examined the effect of sustained, intermittent or cyclic hypoxia on bone health and hormonal responses in humans. Different modes of hypoxic conditioning may have different impacts on bone metabolism both in vivo and in vitro. Additional research is necessary to establish the optimal cyclical dose of oxygen concentration and exposure time.

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