The Impact of Tumor Nitric Oxide Production on VEGFA Expression and Tumor Growth in a Zebrafish Rat Glioma Xenograft Model

Abstract

To investigate the effect of nitric oxide on tumor development, we established a rat tumor xenograft model in zebrafish embryos. The injected tumor cells formed masses in which nitric oxide production could be detected by the use of the cell-permeant DAF-FM-DA (diaminofluorophore 4-amino-5-methylamino-2’-7’-difluorofluorescein diacetate) and DAR-4M-AM (diaminorhodamine-4M). This method revealed that nitric oxide production could be co-localized with the tumor xenograft in 46% of the embryos. In 85% of these embryos, tumors were vascularized and blood vessels were observed on day 4 post injection. Furthermore, we demonstrated by qRT-PCR that the transplanted glioma cells highly expressed Nos2, Vegfa and Cyclin D1 mRNA. In the xenografted embryos we also found increased zebrafish vegfa expression. Glioma and zebrafish derived Vegfa and tumor Cyclin D1 expression could be down regulated by the nitric oxide scavenger 2-(4-Carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide or CPTIO. We conclude that even if there is a heterogeneous nitric oxide production by the xenografted glioma cells that impacts Vegfa and Cyclin D1 expression levels, our results suggest that reduction of nitric oxide levels by nitric oxide scavenging could be an efficient approach to treat glioma.

A novel lncRNA‐LINC01116 regulates tumorigenesis of glioma by targeting VEGFA

“…As a key angiogenic factor, vascular endothelial growth factor (VEGF, also called VEGFA) originally isolated from tumor cells, is involved in glioma cell proliferation, angiogenesis and metastasis. Furthermore, VEGFA exerts its oncogene function may partly through its ability to promote angiogenesis . Although, angiogenesis has been shown to be important in the tumorigenesis and progression of glioma, the effects of angiogenesis on the treatment of tumors are still debated .…”

“…Furthermore, VEGFA exerts its oncogene function may partly through its ability to promote angiogenesis. [11][12][13][14] Although, angiogenesis has been shown to be important in the tumorigenesis and progression of glioma, the effects of angiogenesis on the treatment of tumors are still debated. [15][16][17][18] The multiple functions of VEGFA in the tumorigenesis of glioma need a better understanding of the underlying molecular mechanisms for glioma treatment.…”

Glyceryl trinitrate‑induced cytotoxicity of docetaxel‑resistant prostatic cancer cells is associated with differential regulation of clusterin

“…In the present study a transgenic zebrafish line fli1a: Enhanced Green Fluorescent Protein (EGFP) expressing EGFP in endothelial cells under the fli1a promoter was used (Zebrafish International Resource Center, University of Oregon, Eugene, OR, USA). This model allows the following of eventual neovascularization at the yolksac level following a microinjection of tumor cells (41). Adult zebrafish (15 males and 15 females) were maintained in a recirculating aquaculture system (Müller & Pfleger GmbH & Co. KG, Rockenhausen, Germany) with a temperature range of 26-28˚C, and a 14/10-h light/dark cycle as previously described (41).…”

“…This model allows the following of eventual neovascularization at the yolksac level following a microinjection of tumor cells (41). Adult zebrafish (15 males and 15 females) were maintained in a recirculating aquaculture system (Müller & Pfleger GmbH & Co. KG, Rockenhausen, Germany) with a temperature range of 26-28˚C, and a 14/10-h light/dark cycle as previously described (41). They were fed twice a day with dried flake food.…”

Ti-SLActive and TiZr-SLActive Dental Implant Surfaces Promote Fast Osteoblast Differentiation

“…DAF-FM diacetate and DHR stainings were employed for detection of NO and ROS/RNS [60][61][62][63]. Osteoblasts were cultivated on six surfaces of different topographies and intracellular levels of NO and ROS/RNS were examined.…”