Evaluation of deacetylase inhibition in metaplastic breast carcinoma using multiple derivations of preclinical models of a new patient-derived tumor

AbstractMetaplastic breast carcinoma (MBC) is a clinically aggressive and rare subtype of breast cancer, with similar features to basal-like breast cancers. Due rapid growth rates and characteristic heterogeneity, MBC is often unresponsive to standard chemotherapies; and novel targeted therapeutic discovery is urgently needed. Histone deacetylase inhibitors (DACi) suppress tumor growth and metastasis through regulation of the epithelial-to-mesenchymal transition axis in various cancers, including basal-like breast cancers.We utilized a new MBC patient-derived xenograft (PDX) to examine the effect of DACi therapy on MBC. Cell morphology, cell cycle-associated gene expressions, transwell migration, and metastasis were evaluated in patient-derived cells and tumors after treatment with romidepsin and panobinostat. Derivations of our PDX model, including cells, spheres, organoids, explants, and in vivo implanted tumors were treated. Finally, we tested the effects of combining DACi with approved chemotherapeutics on relative cell biomass.DACi significantly suppressed the total number of lung metastasis in vivo using our PDX model, suggesting a role for DACi in preventing circulating tumor cells from seeding distal tissue sites. These data were supported by our findings that DACi reduced cell migration, populations, and expression of mesenchymal-associated genes. While DACi treatment did affect cell cycle-regulating genes in vitro, tumor growth was not affected compared to controls. Importantly, gene expression results varied depending on the cellular or tumor system used, emphasizing the importance of using multiple derivations of cancer models in preclinical therapeutic discovery research. Furthermore, DACi sensitized and produced a synergistic effect with approved oncology therapeutics on inherently resistant MBC.This study introduced a role for DACi in suppressing the migratory and mesenchymal phenotype of MBC cells through regulation of the epithelial-mesenchymal transition axis and suppression of the CTC population. Preliminary evidence that DACi treatment in combination with MEK1/2 inhibitors exerts a synergistic effect on MBC cells was also demonstrated.

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The Role of p53-Mediated Signaling in the Therapeutic Response of Colorectal Cancer to 9F, a Spermine-Modified Naphthalene Diimide Derivative

Cancers ◽

10.3390/cancers12030528 ◽

2020 ◽

Vol 12 (3) ◽

pp. 528 ◽

Cited By ~ 1

Author(s):

Lei Gao ◽

Chaochao Ge ◽

Senzhen Wang ◽

Xiaojuan Xu ◽

Yongli Feng ◽

...

Keyword(s):

Colorectal Cancer ◽

Cell Cycle ◽

Tumor Growth ◽

Molecular Mechanisms ◽

High Rate ◽

Anticancer Activities ◽

Altered Expression ◽

Mesenchymal Transition

Colorectal cancer (CRC) is one of the most prevalent cancers due to its frequency and high rate of mortality. Polyamine-vectorized anticancer drugs possess multiple biological properties. Of these drugs, 9F has been shown to inhibit tumor growth and the metastasis of hepatocellular carcinoma. This current study aims to investigate the effects of 9F on CRC and determine its molecular mechanisms of action. Our findings demonstrate that 9F inhibits CRC cell growth by inducing apoptosis and cell cycle arrest, and suppresses migration, invasion and angiogenesis in vitro, resulting in the inhibition of tumor growth and metastasis in vivo. Based on RNA-seq data, further bioinformatic analyses suggest that 9F exerts its anticancer activities through p53 signaling, which is responsible for the altered expression of key regulators of the cell cycle, apoptosis, the epithelial-to-mesenchymal transition (EMT), and angiogenesis. In addition, 9F is more effective than amonafide against CRC. These results show that 9F can be considered as a potential strategy for CRC treatment.

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FOXO3a shRNA and simultaneous induction of P27Kip1gene Inhibit the Breast Cancer Growth in Nude Mice

10.21203/rs.3.rs-31573/v1 ◽

2020 ◽

Author(s):

Sabah Mayahi ◽

Masood Golalipour ◽

Mohammad Ashari ◽

Majid Shahbazi

Keyword(s):

Breast Cancer ◽

Cell Cycle ◽

Tumor Growth ◽

Nude Mice ◽

Cell Apoptosis ◽

Immunohistochemical Analysis ◽

Tumor Growth Inhibition ◽

Breast Cancers ◽

Simultaneous Induction

Abstract Background: FOXO proteins, which are overexpressed in multiple human tumors, belong to the Forkhead family of transcription factors that are involved in cell-cycle regulation, cell apoptosis, differentiation, stress response, and metabolism. The p27Kip1 gene leads to cell cycle arrest, cell apoptosis, tumor suppressor genes, and cell adhesion. The low expression level of the p27Kip1 gene is attributed to poor prognosis in patients with colorectal, gastric, pulmonary, and breast cancers. Accordingly, the present study aimed to investigate the possibility of tumor growth inhibition in a mouse model by targeting FOXO3a shRNA and the simultaneous induction of P27Kip1gene.Methods: The tumor model was generated by intratumoral inoculating with plasmids. When tumor size reached an average volume of 8 mm in diameter, the mice received injections of construct and control plasmids three times a week for two weeks, followed by tumor growth assessment.Results: Based on the obtained results, the delivery of construct plasmid significantly inhibited tumor growth in nude mice, as compared to the control plasmid. Moreover, the immunohistochemical analysis indicated that the delivery of construct plasmid significantly suppressed expression of FOXo3a and induced P27Kip1 in tumor samples.Conclusion: The findings of the present study revealed that FOXO3a shRNA, along with simultaneous induction of P27Kip1gene using a useful in vivo gene delivery strategy, seems a practical therapeutic approach for breast cancer treatment and may provide profound insight into gene therapy of solid cancers.

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HDAC4 promotes nasopharyngeal carcinoma progression and serves as a therapeutic target

Cell Death and Disease ◽

10.1038/s41419-021-03417-0 ◽

2021 ◽

Vol 12 (2) ◽

Author(s):

Chun Cheng ◽

Jun Yang ◽

Si-Wei Li ◽

Guofu Huang ◽

Chenxi Li ◽

...

Keyword(s):

Nasopharyngeal Carcinoma ◽

Tumor Growth ◽

Therapeutic Target ◽

Histone Deacetylases ◽

Migration And Invasion ◽

Poor Overall Survival ◽

Mesenchymal Transition ◽

E Cadherin

AbstractHistone deacetylases (HDACs) are involved in tumor progression, and some have been successfully targeted for cancer therapy. The expression of histone deacetylase 4 (HDAC4), a class IIa HDAC, was upregulated in our previous microarray screen. However, the role of HDAC4 dysregulation and mechanisms underlying tumor growth and metastasis in nasopharyngeal carcinoma (NPC) remain elusive. Here, we first confirmed that the HDAC4 levels in primary and metastatic NPC tissues were significantly increased compared with those in normal nasopharyngeal epithelial tissues and found that high HDAC4 expression predicted a poor overall survival (OS) and progression-free survival (PFS). Functionally, HDAC4 accelerated cell cycle G1/S transition and induced the epithelial-to-mesenchymal transition to promote NPC cell proliferation, migration, and invasion in vitro, as well as tumor growth and lung metastasis in vivo. Intriguingly, knockdown of N-CoR abolished the effects of HDAC4 on the invasion and migration abilities of NPC cells. Mechanistically, HDAC3/4 binds to the E-cadherin promoter to repress E-cadherin transcription. We also showed that the HDAC4 inhibitor tasquinimod suppresses tumor growth in NPC. Thus, HDAC4 may be a potential diagnostic marker and therapeutic target in patients with NPC.

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Cancer-associated fibroblast secretion of PDGFC promotes gastrointestinal stromal tumor growth and metastasis

Oncogene ◽

10.1038/s41388-021-01685-w ◽

2021 ◽

Vol 40 (11) ◽

pp. 1957-1973

Author(s):

Hyunho Yoon ◽

Chih-Min Tang ◽

Sudeep Banerjee ◽

Mayra Yebra ◽

Sangkyu Noh ◽

...

Keyword(s):

Signal Transduction ◽

Tumor Growth ◽

Gastrointestinal Stromal Tumor ◽

Single Agent ◽

Stromal Tumor ◽

Paracrine Signaling ◽

Mesenchymal Transition ◽

Tumor Growth And Metastasis ◽

Factor C

AbstractTargeted therapies for gastrointestinal stromal tumor (GIST) are modestly effective, but GIST cannot be cured with single agent tyrosine kinase inhibitors. In this study, we sought to identify new therapeutic targets in GIST by investigating the tumor microenvironment. Here, we identified a paracrine signaling network by which cancer-associated fibroblasts (CAFs) drive GIST growth and metastasis. Specifically, CAFs isolated from human tumors were found to produce high levels of platelet-derived growth factor C (PDGFC), which activated PDGFC-PDGFRA signal transduction in GIST cells that regulated the expression of SLUG, an epithelial-mesenchymal transition (EMT) transcription factor and downstream target of PDGFRA signaling. Together, this paracrine induce signal transduction cascade promoted tumor growth and metastasis in vivo. Moreover, in metastatic GIST patients, SLUG expression positively correlated with tumor size and mitotic index. Given that CAF paracrine signaling modulated GIST biology, we directly targeted CAFs with a dual PI3K/mTOR inhibitor, which synergized with imatinib to increase tumor cell killing and in vivo disease response. Taken together, we identified a previously unappreciated cellular target for GIST therapy in order to improve disease control and cure rates.

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O80: GREMLIN-1 PROMOTES TUMOURIGENESIS AND METASTASIS IN HER2 POSITIVE BREAST CANCER

British Journal of Surgery ◽

10.1093/bjs/znab117.080 ◽

2021 ◽

Vol 108 (Supplement_1) ◽

Author(s):

C Zabkiewicz ◽

L Ye ◽

R Hargest

Keyword(s):

Breast Cancer ◽

Cellular Growth ◽

Breast Cancers ◽

Her2 Positive ◽

Mesenchymal Transition ◽

Her2 Breast Cancer ◽

Bmp Signalling ◽

Gremlin 1

Abstract Introduction HER2 over-expression denotes poor prognosis in breast cancers.Bone morphogenetic protein(BMP) signalling is known to interact with EGF signalling, co-regulating breast cancer progression.BMP antagonist Gremlin-1 may influence breast cancer disease progression, but this remains unexplored in HER2 positive breast cancers. Method GREM1 and HER2 expression, and clinical outcomes were examined in clinical cohorts.GREM1 overexpression or pEF control plasmid were transduced into BT474 HER2+breast cancer cells. In vitro function tests using BT474 pEF and BT474GREM1cells include 2D/3D growth, migration, and expression of epithelial to mesenchymal transition(EMT)markers. Signalling cascades were examined in BT474 treated with RhGremlin-1. In vivo, BALB/c nude mice underwent either mammary injection or intra-cardiac injection of BT474pEF or BT474GREM1 cells and disease burden assessed. Result GREM1 expression correlates with HER2 in breast tumours(p=0.03) and is higher in metastatic HER2 positive cancers (p = 0.04). HER2 positive patients with high GREM1 have poor survival(p = 0.0002). BT474GREM1cells have up-regulated markers of EMT compared to control. BT474 RhGremlin-1 treated cells have active AKT pathway signalling, independent of BMP signalling. In vitro, BT474GREM1cells significantly proliferate and migrate compared to control(p<0.05 and p < 0.001).This is confirmed in vivo, BT474GREM1 mice grew significantly larger mammary tumours(p<0.05) and had more PETCT metastatic hotspots. Conclusion Gremlin-1 is correlated with poor outcomes in HER2 patients and promotes breast cancer cellular growth, migration and metastasis.Gremlin-1 is a novel area of research with potential as a prognostic biomarker and therapeutic target for personalised, effective, breast cancer outcomes. Take-home message BMP antagonists are gaining interest for their potential in breast cancer prognosis and therapeutics.This novel area of research shows BMP antagonist Gremlin-1 is of importance in HER2 positive breast cancers. DRAGONS DEN

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A Novel Bispecific Antibody Targeting EGFR and VEGFR2 Is Effective against Triple Negative Breast Cancer via Multiple Mechanisms of Action

Cancers ◽

10.3390/cancers13051027 ◽

2021 ◽

Vol 13 (5) ◽

pp. 1027

Author(s):

Nishant Mohan ◽

Xiao Luo ◽

Yi Shen ◽

Zachary Olson ◽

Atul Agrawal ◽

...

Keyword(s):

Endothelial Cells ◽

Tumor Growth ◽

Mechanisms Of Action ◽

Breast Cancers ◽

Single Chain ◽

Cancer Cell Growth ◽

Anti Tumor Activity ◽

Enhance Tumor Growth

Both EGFR and VEGFR2 frequently overexpress in TNBC and cooperate with each other in autocrine and paracrine manner to enhance tumor growth and angiogenesis. Therapeutic mAbs targeting EGFR (cetuximab) and VEGFR2 (ramucirumab) are approved by FDA for numerous cancer indications, but none of them are approved to treat breast cancers. TNBC cells secrete VEGF-A, which mediates angiogenesis on endothelial cells in a paracrine fashion, as well as promotes cancer cell growth in autocrine manner. To disrupt autocrine/paracrine loop in TNBC models in addition to mediating anti-EGFR tumor growth signaling and anti-VEGFR2 angiogenic pathway, we generated a BsAb co-targeting EGFR and VEGFR2 (designated as anti-EGFR/VEGFR2 BsAb), using publicly available sequences in which cetuximab IgG backbone is connected to the single chain variable fragment (scFv) of ramucirumab via a glycine linker. Physiochemical characterization data shows that anti-EGFR/VEGFR2 BsAb binds to both EGFR and VEGFR2 in a similar binding affinity comparable to parental antibodies. Anti-EGFR/VEGFR2 BsAb demonstrates in vitro and in vivo anti-tumor activity in TNBC models. Mechanistically, anti-EGFR/VEGFR2 BsAb not only directly inhibits both EGFR and VEGFR2 in TNBC cells but also disrupts autocrine mechanism in TNBC xenograft mouse model. Furthermore, anti-EGFR/VEGFR2 BsAb inhibits ligand-induced activation of VEGFR2 and blocks paracrine pathway mediated by VEGF secreted from TNBC cells in endothelial cells. Collectively, our novel findings demonstrate that anti-EGFR/VEGFR2 BsAb inhibits tumor growth via multiple mechanisms of action and warrants further investigation as a targeted antibody therapeutic for the treatment of TNBC.

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Spindle Cell Metaplastic Breast Carcinoma

Current Medical Imaging Formerly Current Medical Imaging Reviews ◽

10.2174/1573405617666211004114041 ◽

2021 ◽

Vol 17 ◽

Author(s):

Vithya Sanmugasiva ◽

Marlina Tanty Ramli Hamid ◽

Farhana Fadzli ◽

Nazimah Ab Mumin ◽

Kartini Rahmat

Keyword(s):

Breast Carcinoma ◽

Cell Carcinoma ◽

Spindle Cell ◽

Final Diagnosis ◽

Spindle Cell Carcinoma ◽

Left Breast ◽

Metaplastic Carcinoma ◽

Breast Cancers ◽

Breast Lump ◽

Metaplastic Breast Carcinoma

Introduction: Metaplastic breast carcinoma is an uncommon malignancy that constitutes < 5% of all breast cancers. There are 5 subtypes which are spindle cell, squamous cell, carcinosarcoma, matrix-producing and metaplastic with osteoclastic giant cells. Spindle cell carcinoma represents approximately <0.3% of invasive breast carcinomas. It is typically a triple-negative cancer with distinct pathological characteristics, but relatively a non-conclusive imaging findings. Case report: An elderly lady presented with an enlarging painful left breast lump for 1 year. Palpable left breast lump noted on clinical examination. Mammography demonstrated a high density, oval lesion with a partially indistinct margin. Corresponding ultrasound showed a large irregular heterogeneous lesion with solid-cystic areas. Histopathology showed atypical spindle-shaped cells which stained positive for cytokeratins and negative for hormone and human epidermal growth factor receptors, which favours spindle cell metaplastic carcinoma. Left mastectomy and axillary dissection were performed, and the final diagnosis was consistent with metaplastic spindle cell carcinoma. Conclusion: Spindle cell carcinoma of the breast is a rare aggressive histological type of carcinoma which may present with benign features on imaging. Tissue diagnosis is essential for prompt diagnosis with multidisciplinary team discussion to guide management and improve patient’s outcome.

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LncRNA SNHG12 regulates the radiosensitivity of cervical cancer through the miR-148a/CDK1 pathway

Cancer Cell International ◽

10.1186/s12935-020-01654-5 ◽

2020 ◽

Vol 20 (1) ◽

Author(s):

Chen Wang ◽

Shiqing Shao ◽

Li Deng ◽

Shelian Wang ◽

Yongyan Zhang

Keyword(s):

Cell Cycle ◽

Cervical Cancer ◽

Tumor Growth ◽

Luciferase Reporter ◽

Tunel Staining ◽

Cell Cycle Distribution ◽

Ki 67 ◽

In Vivo Experiments ◽

Potential Biomarker

Abstract Background Radiation resistance is a major obstacle to the prognosis of cervical cancer (CC) patients. Many studies have confirmed that long non-coding RNAs (lncRNAs) are involved in the regulation of radiosensitivity of cancers. However, whether small nucleolar RNA host gene 12 (SNHG12) regulates the radiosensitivity of CC remains unknown. Methods Quantitative real-time polymerase chain reaction was used to measure the expression levels of SNHG12 and microRNA-148a (miR-148a). The radiosensitivity of cells was evaluated by clonogenic assay. Flow cytometry and caspase-3 activity assay were performed to assess the apoptosis ability and cell cycle distribution of cells. Besides, dual-luciferase reporter and RNA immunoprecipitation assay were used to verify the interaction between miR-148a and SNHG12 or cyclin-dependent kinase 1 (CDK1). Also, the protein levels of CDK1, CCND1 and γ-H2AX were detected by western blot analysis. Furthermore, in vivo experiments were conducted to verify the effect of SNHG12 on CC tumor growth. Ki-67 and TUNEL staining were employed to evaluate the proliferation and apoptosis rates in vivo. The hematoxylin and eosin (HE) staining were employed to evaluate the tumor cell morphology. Results SNHG12 was upregulated in CC tissues and cells, and its knockdown improved the radiosensitivity by promoting the radiation-induced apoptosis and cell cycle arrest of CC cells. Also, miR-148a could be sponged by SNHG12 and could target CDK1. MiR-148a inhibitor or CDK1 overexpression could invert the promotion effect of silenced-SNHG12 on CC radiosensitivity. Meanwhile, SNHG12 interference reduced the tumor growth of CC, increased miR-148a expression, and inhibited CDK1 level in vivo. Conclusion LncRNA SNHG12 promoted CDK1 expression to regulate the sensitivity of CC cells to radiation through sponging miR-148a, indicating that SNHG12 could be used as a potential biomarker to treat the radiotherapy resistance of CC patients.

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LncRNA MONC Suppresses the Malignant Phenotype of Endometrial Cancer Stem Cells by Regulating the MiR-636/GLCE Axis

10.21203/rs.3.rs-40559/v1 ◽

2020 ◽

Author(s):

Yibing Li ◽

Jianing Huo ◽

Junjian He ◽

Haining Ma ◽

Xiaoxin Ma

Keyword(s):

Cell Cycle ◽

Stem Cells ◽

Endometrial Cancer ◽

Cancer Stem Cells ◽

Suppressive Effect ◽

Luciferase Assay ◽

Biological Behavior ◽

Mesenchymal Transition ◽

Tumor Suppressive Effect

Abstract Background: Emerging evidence shows that abnormal expression of long non-coding RNA is involved in the occurrence and development of various tumors. LncRNA MONC is abnormally expressed in head and neck squamous cell carcinoma (HNSCC), lung cancer, colorectal cancer, and acute megakaryocytic leukemia, but the biological function and potential regulatory mechanism of MONC in endometrial cancer stem cells (ECSCs) and endometrial cancer cells (ECCs) have not been studied. In this study, we aimed to explore the tumor suppressive effect and mechanism of MONC in regulating ECSCs and ECCs. Methods: The expression of genes was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The expression of proteins was detected by Western blot. The interplay of LncRNA-miRNA-mRNA was verified using the luciferase assay. The growth rate of ECSC spheroids was detected by sphere formation assay. Cell proliferation was detected by CCK-8 assay. The cell invasion was detected by transwell invasion assay. Cell cycle was detected by Cell cycle analysis.Cell apoptosis was detected by the Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) double-staining assay. Animal study was conducted to evaluate the effect of MONC combined with miR-636 on tumor growth in vivo. Results: Low MONC expression in endometrial carcinoma (EC), which directly inhibits the malignant biological behavior of ECSCs and ECCs by directly inhibiting miR-636. Simultaneously, miR-636 may indirectly reduce the expression of MONC. Down-regulation of miR-636 may promote GLCE expression by targeting the 3'-untranslated region (UTR) of the downstream gene GLCE, thereby inhibiting the progression of ECSCs. MONC combined with miR-636 inhibited the Notch signaling pathway and tumor epithelial-to-mesenchymal transition (EMT) process. In addition, we verified the tumor suppressive effect of MONC in nude mice, miR-636 can rescue the tumor suppressive effect of overexpressing MONC, and this effect is more obvious in ECSC. Conclusion: MONC inhibits the malignant phenotypes of ECSCs and ECCs by regulating the miR-636/GLCE axis. The MONC/miR-636/GLCE axis may provide novel treatment avenues for human EC.

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RUNX1 suppresses breast cancer stemness and tumor growth

10.1101/315093 ◽

2018 ◽

Author(s):

Deli Hong ◽

Andrew J. Fritz ◽

Kristiaan H. Finstad ◽

Mark P. Fitzgerald ◽

Adam L. Viens ◽

...

Keyword(s):

Breast Cancer ◽

Stem Cell ◽

Tumor Suppressor ◽

Cancer Stem Cell ◽

Tumor Growth ◽

Epithelial Mesenchymal Transition ◽

Ectopic Expression ◽

Cell Activity ◽

Mesenchymal Transition

SummaryRecent studies have revealed that mutations in the transcription factor Runx1 are prevalent in breast tumors. Yet, how loss of Runx1 contributes to breast cancer (BCa) remains unresolved. We demonstrate for the first time that Runx1 represses the breast cancer stem cell (BCSC) phenotype and consequently, functions as a tumor suppressor in breast cancer. Runx1 ectopic expression in MCF10AT1 and MCF10CA1a BCa cells reduces (60%) migration, invasion and in vivo tumor growth in mouse mammary fat pad (P<0.05). Runx1 is decreased in BCSCs, and overexpression of Runx1 suppresses tumorsphere formation and reduces the BCSC population. Furthermore, Runx1 inhibits Zeb1 expression, while Runx1 depletion activates Zeb1 and the epithelial-mesenchymal transition. Mechanistically Runx1 functions as a tumor suppressor in breast cancer through repression of cancer stem cell activity. This key regulation of BCSCs by Runx1 may be shared in other epithelial carcinomas, highlighting the importance of Runx1 in solid tumors.

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