The Histone Methyltransferase EZH2 Mediates Tumor Progression on the Chick Chorioallantoic Membrane Assay, a Novel Model of Head and Neck Squamous Cell Carcinoma

Liu, Min; Scanlon, Christina Springstead; Banerjee, Rajat; Russo, Nickole; Inglehart, Ronald; Willis, Amanda L.; Weiss, Stephen J.; D’Silva, Nisha J.

doi:10.1593/tlo.13175

Cited by 58 publications

(73 citation statements)

References 34 publications

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“…Advantages are that the CAM model lacks immunological and inflammatory cells, making it an attractive system to avoid effects of the tumor microenvironment interfering with tumor development [6]. This has been used to provide highly reproducible models for glioblastoma [7,8], bone and osteosarcoma [5,9], pancreatic adenocarcinoma [10], ovarian cancer [11], head and neck squamous carcinoma [12], and more recently renal carcinoma [13], as well as CRC [14]. In the last case, the development of CRC and therapeutic escape may be caused by the presence of tumor-initiating cells (TICs) [3,15], which are responsible for driving growth and treatment resistance [16,17].…”

Section: Introductionmentioning

confidence: 99%

New ex-ovo colorectal-cancer models from different SdFFF-sorted tumor-initiating cells

et al. 2015

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Despite effective treatments, relapse of colorectal cancer (CRC) is frequent, in part caused by the existence of tumor-initiating cells (TICs). Different subtypes of TICs, quiescent and activated, coexist in tumors, defining the tumor aggressiveness and therapeutic response. These subtypes have been sorted by hyperlayer sedimentation field-flow fractionation (SdFFF) from WiDr and HCT116 cell lines. On the basis of a new strategy, including TIC SdFFF sorting, 3D Matrigel amplification, and grafting of corresponding TIC colonies on the chick chorioallantoic membrane (CAM), specific tumor matrices could be obtained. If tumors had similar architectural structure with vascularization by the host system, they had different proliferative indices in agreement with their initial quiescent or activated state. Protein analysis also revealed that tumors obtained from a population enriched for "activated" TICs lost "stemness" properties and became invasive. In contrast, tumors obtained from a population enriched for "quiescent" TICs kept their stemness properties and seemed to be less proliferative and invasive. Then, it was possible to produce different kinds of tumor which could be used as selective supports to study carcinogenesis and therapy sensitivity.

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Section: Introductionmentioning

confidence: 99%

New ex-ovo colorectal-cancer models from different SdFFF-sorted tumor-initiating cells

et al. 2015

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“…The chicken chorioallantoic membrane (CAM) assay is an established in vivo model for investigating the process of new blood vessel formation and vessel responses to anti-angiogenic agents (13). We thereby applied the CAM assay to assess the effect of rH on angiogenesis.…”

Section: Methodsmentioning

confidence: 99%

Recombinant hirudin suppresses the viability, adhesion, migration and invasion of Hep-2 human laryngeal cancer cells

Lü

et al. 2015

Oncology Reports

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Abstract. Recombinant hirudin (rH) is a highly potent and specific inhibitor of thrombin, and has been shown to inhibit the growth and metastasis of several types of cancers in experimental tumor models. The objective of this study was to evaluate the antitumor effects and explore the underlying mechanisms of rH in Hep-2 human laryngeal carcinoma (LC) cells. Hep-2 cells were treated with various concentrations of rH for 24 h. The cell viability was evaluated by a water-soluble tetrazolium salt (WST) assay. The adhesion ability of the cells was evaluated by cell adhesion to fibronectin. Cell migration and invasion were measured with the Boyden chamber assay. Cell apoptosis was detected by Hoechst 33324 fluorescence staining. A chicken chorioallantoic membrane (CAM) assay was used to assess the effects of rH on angiogenesis in vivo. Western blotting was used to detect the expression levels of vascular endothelial growth factor receptor (VEGF-R), focal adhesion kinase (FAK), Bcl-2-associated agonist of cell death (Bad) and B-cell CLL/lymphoma 2 (Bcl-2) proteins. rH significantly inhibited the cell viability and induced apoptosis in LC Hep-2 cells in a dose-dependent manner, as compared with phosphate-buffered saline (PBS) as control. These results were accompanied by a decrease in the anti-apoptotic protein Bcl-2 and an increase in the pro-apoptotic protein Bad. Moreover, rH dose-dependently inhibited the adhesion, migration and invasion of the Hep-2 cells, compared to the vehicle PBS. In addition, rH robustly suppressed angiogenesis in the CAM assay. Importantly, the expression of adhesion and angiogenesis-associated proteins FAK and VEGF-R was significantly downregulated by rH in a dose-dependent manner. The present findings demonstrate that rH exerts antitumor effects in Hep-2 human laryngeal cancer cells via multiple mechanisms and suggests that targeting thrombin by rH is a potential strategy for the treatment of LC. IntroductionLaryngeal carcinoma (LC) is a common head and neck malignancy and accounts for ~2.4% of newly diagnosed malignancies worldwide every year (1,2). Surgery, chemotherapy and radiation therapy are the current treatment modules for laryngeal cancer (3-5). Although early-stage laryngeal cancer can be effectively treated with surgery or radiotherapy, the 5-year survival rate of patients with advanced LC is still below 60% even after systematic surgery and post-surgical adjuvant radiotherapy or chemotherapy (6). In addition, surgery may result in complete or partial loss of swallowing and vocal functions. Many patients have to maintain a tracheal cannula on a long-term basis due to laryngeal stenosis after surgery, which markedly impairs their quality of life (6,7). Therefore, there is urgent need to develop novel approaches and strategies for the treatment of these advanced LC patients.The development and progression of cancer are closely associated with thrombosis (8). Thrombin, the key terminal enzyme of coagulation, enhances angiogenesis, stimulates the adhesion of tumor cells to platelet ...

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“…In sub-cutaneous or sub-mucosal injections, in vitro cultured tumor cells are injected in the back or hind-leg of the mouse to produce tumors at the injection site. Since tumor cells are injected directly into the connective tissue, bypassing the basement membrane, the crucial early steps of invasion that transform a pre-cancerous lesion to HNSCC cannot be investigated [9, 50]. However, these models are used to investigate other oncogenic phenotypes including angiogenesis, metastasis and the impact of proteins or other chemicals on tumor size [51–53].…”

Section: In Vivo Assaysmentioning

confidence: 99%

“…The CAM has been used to study angiogenesis and metastasis [63, 64]. Studying HNSCC invasion with CAM is a recent innovation [50]. …”

Section: In Vivo Assaysmentioning

confidence: 99%

“…This membrane is separated from the CAM, which is “dropped” with Hank’s buffered salt solution (HBSS) and suction [50]. Subsequently, the window is enlarged and the outer membrane removed to allow HNSCC cells to be seeded on the CAM [50]. Finally, the window is covered to prevent desiccation and the egg is incubated for multiple days for tumor development on the CAM.…”

Section: In Vivo Assaysmentioning

confidence: 99%

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Reviewing and reconsidering invasion assays in head and neck cancer

2014

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Head and neck squamous cell carcinomas (HNSCC) are malignant tumors that arise from the surface epithelium of the oral cavity, oropharynx and larynx, primarily due to exposure to chemical carcinogens or the human papilloma virus. Due to their location, dental practitioners are well-positioned to detect the lesions. Deadlier than lymphoma or melanoma, HNSCC is incompletely understood. For these reasons, dental practitioners and researchers are focused on understanding HNSCC and the processes driving it. One of these critical processes is invasion, the degradation of the basement membrane by HNSCC cells with subsequent movement into the underlying connective tissue, blood vessels or nerves. Cancer cells metastasize to distant sites via the blood vessels, lymphatics and nerves. Metastasis is associated with poor survival. Since invasion is essential for development and metastasis of HNSCC, it is essential to understand the mechanism(s) driving this process. Elucidation of the mechanisms involved will facilitate the development of targeted treatment, thereby accelerating development of precision/ personalized medicine to treat HNSCC. Robust in vitro and in vivo assays are required to investigate the mechanistic basis of invasion. This review will focus on in vitro and in vivo assays used to study invasion in HNSCC, with special emphasis on some of the latest assays to study HNSCC.

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The Histone Methyltransferase EZH2 Mediates Tumor Progression on the Chick Chorioallantoic Membrane Assay, a Novel Model of Head and Neck Squamous Cell Carcinoma

Cited by 58 publications

References 34 publications

New ex-ovo colorectal-cancer models from different SdFFF-sorted tumor-initiating cells

New ex-ovo colorectal-cancer models from different SdFFF-sorted tumor-initiating cells

Recombinant hirudin suppresses the viability, adhesion, migration and invasion of Hep-2 human laryngeal cancer cells

Reviewing and reconsidering invasion assays in head and neck cancer

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