Advanced tube formation assay using human endothelial colony forming cells for <i>in vitro</i> evaluation of angiogenesis

Lee, Hyun‐Sook; Kang, Kyu‐Tae

doi:10.4196/kjpp.2018.22.6.705

Cited by 18 publications

(12 citation statements)

References 14 publications

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“…However, it should be highlighted that this beneficial effect was mainly based on the demonstration that ACE-Is or ARBs inhibit the process of angiogenesis, the most studied and extensively exploited biological mechanism. The main aspects explored included: the involvement of VEGF serum levels or VEGF protein or mRNA expression[25,28-30,33-35,37-42] in endothelial cell growth and differentiation[46]; the contribution of HIF-1[28,40] as a transcription factor for VEGF[47]; the use of CD31[29-33,42] [also known as platelet endothelial cell adhesion molecule 1 (PECAM-1)] to recognize endothelial cells[48]; endothelial cell proliferation and tubular formation in vitro and/or in vivo [30,32,34,37,39,41,42], a multi-step process triggered by VEGF[49].…”

Section: Discussionmentioning

confidence: 99%

Systematic review: Renin-angiotensin system inhibitors in chemoprevention of hepatocellular carcinoma

Barone¹,

Viggiani²,

Losurdo³

et al. 2019

WJG

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BACKGROUND Neoangiogenesis is one of the key pathogenetic mechanisms in hepatocellular carcinoma (HCC). Modulation of the renin-angiotensin system (RAS) by angiotensin-converting enzyme inhibitors (ACE-Is) and angiotensin receptor blockers (ARBs) seems to be a possible adjuvant therapy for HCC, due to the anti-angiogenic and anti-fibrogenic activity of these drugs. AIM To elucidate the role of ARBs and ACE-Is in HCC. METHODS We performed an electronic search of the literature using the most accessed online databases (PubMed, Cochrane library, Scopus and Web of Science), entering the query terms "angiotensin-converting enzyme inhibitors" OR "ACE inhibitors" OR "ACE-I" AND "hepatocarcinoma*" OR "hepatocellular carcinoma; moreover "angiotensin II type 1 receptor blockers" OR "ARBs" AND "hepatocarcinoma*" OR "hepatocellular carcinoma". Eligibility criteria were: (1) prospective or retrospective clinical studies; (2) epidemiological studies; and (3) experimental studies conducted in vivo or in vitro . Abstracts, conference papers, and reviews were excluded a priori. We limited our literature search to articles published in English, in peer-reviewed journals. RESULTS Thirty-one studies were selected. Three interventional studies showed that ACE-Is had a significant protective effect on HCC recurrence only when used in combination with vitamin K or branched chain aminoacids, without a significant increase in overall survival. Of six retrospective observational studies, mainly focused on overall survival, only one demonstrated a prolonged survival in the ACE-Is group, whereas the two that also evaluated tumor recurrence showed conflicting results. All experimental studies displayed beneficial effects of RAS inhibitors on hepatocarcinogenesis. Numerous experimental studies, conducted either on animals and cell cultures, demonstrated the anti-angiogenetic and antifibrotic effect of ACE-Is and ARBs, thanks to the suppression of some cytokines such as vascular endothelial growth factor, hypoxia-inducible factor-1a, transforming growth factor-beta and tumor necrosis factor alpha. All or parts of these mechanisms were demonstrated in rodents developing fewer HCC and preneoplastic lesions after receiving such drugs. CONCLUSION In humans, RAS inhibitors - alone or in combination - significantly suppressed the cumulative HCC recurrence, without prolonging patient survival, but some limitations intrinsic to these studies prompt further investigations.

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Section: Discussionmentioning

confidence: 99%

Systematic review: Renin-angiotensin system inhibitors in chemoprevention of hepatocellular carcinoma

Barone¹,

Viggiani²,

Losurdo³

et al. 2019

WJG

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“…The capability of ECFCs to form tubule-like structures was determined using a tube formation assay with modifications of previous procedures. 15 The 24-well plates were coated with ice-cold Matrigel solution (Phenol Red-Free; BD Biosciences) and incubated at 37°C for 30 minutes to allow the Matrigel to solidify. ECFCs were harvested, suspended in EBM-MV 2 (2% FBS and 1% GPS) or CM (1:1 dilution with EBM-MV 2 containing 2% FBS and 1% GPS), and added to the Matrigel-coated 24-well plates at 15×10 4 cells/well.…”

Section: Methodsmentioning

confidence: 99%

Differential Angiogenic Responses of Human Endothelial Colony-Forming Cells to Different Molecular Subtypes of Breast Cancer Cells

Lee

Kang

2021

J Lipid Atheroscler

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Objective Triple negative breast cancer (TNBC) is one subtype of breast cancer. It is characterized by lack of estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2. Compared with non-TNBC, TNBC is more aggressive, of higher grade, and frequently metastatic with poor prognosis, which is correlated with upregulated microvascular density. Endothelial colony-forming cells (ECFCs) mediate neovascularization, which is the crucial contributor to cancer growth and metastasis. The present study aimed to determine whether angiogenic responses of ECFCs are regulated differently by TNBC compared with non-TNBC. Methods MDA-MB-231 and MCF7 cells were utilized for TNBC and non-TNBC, respectively. Bone-marrow-derived human ECFCs were treated with a conditioned medium (CM) of cancer cells to investigate the paracrine effect on angiogenesis. Also, ECFCs were co-cultured with cancer cells to evaluate the angiogenic effect of direct cell-to-cell interaction. Angiogenic responses of ECFCs were evaluated by proliferation, migration, and tube formation. Gene expression profiles of pro-angiogenic factors were also analyzed. Results Migration and tube formation of ECFCs were increased by treatment with CM of MDA-MB-231, which correlated with a higher gene expression profile of pro-angiogenic factors in MDA-MB-231 compared to MCF7. Interestingly, ECFCs co-cultured with MDA-MB-231 showed further increase of tube formation, suggesting synergic mechanisms between the paracrine effect and direct interaction between the cells. Conclusion The angiogenic potential of ECFCs was enhanced by TNBC through both direct and indirect mechanisms. Therefore, the investigation of signaling pathways to regulate ECFC-mediated angiogenesis will be important to the discovery of anti-angiogenic therapies to treat TNBC patients.

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“…Matrigel (8.7 mg/mL) was placed in a refrigerator at 4 • C to slowly melt overnight, and 50µL was spread in each well of a 96-well plate and placed in an incubator at 37 • C for 1 h. The EA•hy926 cells were prepared into a cell suspension of 2 × 10 5 cells/mL, which was mixed well and added to a 96-well plate at a volume of 100 µL per well. The CM prepared in advance was then added at a volume of 50 µL per well, incubated in an incubator at 37 • C and 5% CO 2 , and observed after 8 h (Arnaoutova & Kleinman, 2010;Lee & Kang, 2018;Ma et al, 2017). We observed the tube formation of endothelial cells and counted the number of vessels and visible nodes.…”

Section: Vascular Endothelial Cell Tube Formation Experimentsmentioning

confidence: 99%

Tumour-associated macrophages mediate the invasion and metastasis of bladder cancer cells through CXCL8

Zhang

Han

et al. 2020

PeerJ

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Tumour-associated macrophages (TAMs) are associated with both the progression and poor prognosis of a variety of solid tumours. This study aimed to investigate and clarify the tumour-promoting role of CXCL8 secreted by TAMs in the urothelial carcinoma microenvironment of the bladder. Immunohistochemistry (n = 55) was used to detect Chemokine (C-X-C motif) ligand 8 (CXCL8), CD163 (a TAM marker), Matrixmetalloproteinase-9 (MMP-9), vascular endothelial growth factor (VEGF), and E-cadherin in cancerous and adjacent tissues of bladder cancer patients. TAMs-like PBM (peripheral blood mononuclear)-derived macrophages were developed using in vitro experiments. T24, 5637, and UM-UC-3 were treated with conditioned medium (CM) for the experimental intervention group, without CM for the blank control group, and with CM and an anti-CXCL8 neutralizing antibody for the experimental control group, respectively. The immunohistochemical study showed that the expression of CXCL8 was significantly upregulated as the number of infiltrating TAMs increased in the tumour tissues. A high expression of CXCL8 significantly correlated with an increase in the expression of MMP-9 and VEGF and a decrease in expression of E-cadherin in the microenvironment. This revealed that TAM-derived CXCL8 is highly associated with bladder cancer migration, invasion, and angiogenesis. The concentration of CXCL8 was significantly higher in CM collected from TAM-like PBM-derived macrophages than that from THP-1 cells. In subsequent in vitro experiments, we found that CM derived from TAM-like PBM-derived macrophages can also increase the migration rate, invasiveness, and pro-angiogenic properties of tumour cells. Additionally, the effect of CXCL8 was significantly diminished by the addition of an anti-CXCL8 neutralizing antibody to CM. The infiltration of TAMs in the tumour microenvironment leads to the elevation of CXCL8, which in turn promotes the secretion of MMP-9, VEGF, and E-cadherin by bladder cancer cells. This alters the migration, invasion, and pro-angiogenic capacity of bladder cancer cells and accelerates cancer progression.

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Advanced tube formation assay using human endothelial colony forming cells for in vitro evaluation of angiogenesis

Cited by 18 publications

References 14 publications

Systematic review: Renin-angiotensin system inhibitors in chemoprevention of hepatocellular carcinoma

Systematic review: Renin-angiotensin system inhibitors in chemoprevention of hepatocellular carcinoma

Differential Angiogenic Responses of Human Endothelial Colony-Forming Cells to Different Molecular Subtypes of Breast Cancer Cells

Tumour-associated macrophages mediate the invasion and metastasis of bladder cancer cells through CXCL8

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