Comparative proteome analysis of monolayer and spheroid culture of canine osteosarcoma cells

Gebhard, Christiane; Miller, Ingrid; Hummel, Karin Anna; Ondrovics, Martina; Schlosser, Sarah; Walter, Ingrid

doi:10.1016/j.jprot.2018.01.006

Cited by 16 publications

(14 citation statements)

References 70 publications

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“…All of them showed differential expressions between samples of untreated and treated. Most importantly the calcium-binding Protein S100 A4 and A10 showed higher levels of expression in OGS control were reported earlier in patients with metastasis and poor prognosis 30,31 . Interestingly, after freezing and irradiation treatment, both the proteins were reduced.…”

Section: Signaling Proteins Expression In Ogssupporting

confidence: 50%

Proteomic profiling and identification of significant markers from high-grade osteosarcoma after cryotherapy and irradiation

Madda

Chen

Wang

et al. 2020

Sci Rep

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Biological reconstruction of allografts and recycled autografts have been widely implemented in highgrade osteogenic sarcoma. For treating tumor-bearing autografts, extracorporeal irradiation (ECIR) and liquid nitrogen (LN) freezing techniques are being used worldwide as a gold standard treatment procedure. Both the methods aim to eradicate the tumor cells from the local recurrence and restore the limb function. Therefore, it is essential and crucial to find, and compare the alterations at molecular and physiological levels of the treated and untreated OGS recycled autografts to obtain valuable clinical information for better clinical practice. Thus, we aimed to investigate the significantly expressed altered proteins from ECIR-and cryotherapy/freezing-treated OGS (n = 12) were compared to untreated OGS (n = 12) samples using LC-ESI-MS/MS analysis, and the selected proteins from this protein panel were verified using immunoblot analysis. From our comparative proteomic analysis identified a total of 131 differentially expressed proteins (DEPs) from OGS. Among these, 91 proteins were up-regulated (2.5 to 3.5-folds), and 40 proteins were down-regulated (0.2 to 0.5 folds) (p < 0.01 and 0.05). The functional enrichment analysis revealed that the identified DEPs have belonged to more than 10 different protein categories include cytoskeletal, extracellular matrix, immune, enzyme modulators, and cell signaling molecules. Among these, we have confirmed two potential candidates' expressions levels such as Fibronectin and Protein S100 A4 using western blot analysis. Our proteomic study revealed that LN-freezing and ECIR treatments are effectively eradicating tumor cells, and reducing the higher expressions of DEPs at molecular levels which may help in restoring the limb functions of OGS autografts effectively. To the best of our knowledge, this is the first proteomic study that compared proteomic profiles among freezing, ECIR treated with untreated OGS in recycled autografts. Moreover, the verified proteins could be used as prognostic or diagnostic markers that reveal valuable scientific information which may open various therapeutic avenues in clinical practice to improve patient outcomes.High-grade osteogenic sarcomas (OGS) are the most common primary malignant bone sarcomas that distress the bone and forms a matrix and osteoid around the knees 1-3 . It accounts one to three per million each year worldwide and has a high rate of incidence in children and adults 3,4 . Currently, the standard treatment procedures applicable for patients are neoadjuvant chemotherapy drugs combined with surgery, precision diagnostic instruments, and limb salvage operations 5 . At present, there are three reconstructive procedures available after resection of tumors that are affected with major joints, include tumor prosthesis, an osteoarticular allograft, and a composite biological reconstruction. Among these three options, biological reconstruction of allograft and autografts (recycled from the resected autogenic bone segment) technique has ...

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Section: Signaling Proteins Expression In Ogssupporting

confidence: 50%

Proteomic profiling and identification of significant markers from high-grade osteosarcoma after cryotherapy and irradiation

Madda

Chen

Wang

et al. 2020

Sci Rep

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“…It is extremely important to establish an experimental model as close as possible to physiological conditions to enable improvement in the field. It has been previously shown that cells in 3D culture differ in proteomic profile from 2D cultured cells mainly in terms of proteins that are the part of a functional network including structural proteins, chaperones, as well as oxidative stress-related proteins [17]. Moreover, 3D cultured cells respond more effectively than cells from monolayer to cytotoxic agents, such as hydrogen peroxide or heavy metals [11].…”

Section: Discussionmentioning

confidence: 99%

The Differences in the Proteome Profile of Cannabidiol-Treated Skin Fibroblasts following UVA or UVB Irradiation in 2D and 3D Cell Cultures

Gęgotek

Atalay

Domingues

et al. 2019

Cells

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Cannabidiol (CBD), as the only phytocannabinoid that has no psychoactive effect, has both antioxidant and anti-inflammatory effects, and thus might be suggested as a cytoprotective compound against UV-induced metabolic changes in skin cells. Therefore, the aim of this study was to investigate the level of protective CBD activity by evaluating the proteomic profile of 2D and 3D cultured skin fibroblasts models following exposure to UVA and UVB radiation. The CBD cytoprotective effect against UV-induced damage in 2D and 3D cultured fibroblasts were different. The main alterations focus on the range of cell reaction and involved different proteins associated with various molecular functions. In the 2D cultured cells, following UV radiation, the major changes were associated with proteins involved in antioxidant response and inflammation, while, in the 3D cultured fibroblasts, CBD action against UV induced changes were mainly associated with the activation of signalling pathways. Therefore, the knowledge of the CBD action in a multilayer skin cells model allowed for the prediction of changes in cell-cell interactions and skin cell metabolism. Knowledge about the lower protective effect of CBD in 3D cultured fibroblasts should be taken into account during the design of UV light protection.

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“…Chaperon’s family, which is composed by protein folders associated to cellular stress response and cytoskeletal organization, is similarly modulated by 3D context. On the opposite manner, general protein phosphorylation is upregulated in 2D cultures compared to 3D environment, probably due to the increase in the growth rate observed in monolayer cell cultures [86] . These data suggest that, in order to better understand the proteomic profiles presented in tumours, all the previous information obtained by 2D studies must be re-evaluated in the light of the 3D culture methods close to the tumour behaviour.…”

Section: D Culture Methods Of Primary Bone Tumoursmentioning

confidence: 98%

“…Interestingly, the protein expression profiles significantly differ between 2D and 3D cultures. A proteomic study using spheroids produced by ultra-low attachment supports from the dog OS cell line D17 demonstrated that the development in 3D culture induces an increase of glycolysis/glucogenesis pathways, biosynthesis of amino acids and changes in carbohydrate metabolism [86] . These data were in agreement with the metabolism observed during tumour development and the generation of a hypoxic local environment.…”

Section: D Culture Methods Of Primary Bone Tumoursmentioning

confidence: 99%

In vitro three-dimensional cell cultures for bone sarcomas

Muñoz-García

Jubelin

Loussouarn

et al. 2021

Journal of Bone Oncology

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Highlights 3D cell cultures present similar drugs resistance features than in vivo tumours. Unlike 2D cell cultures, 3D culture approaches mimic better the tumour microenvironment. Collagen scaffolds result in a better osteosarcoma 3D proliferation than soft hydrogels. Osteosarcoma 3D models constitute an appealing approach for bone mineralisation studies. Combination of microfluidic/bioprinting technology with bone 3D cultures as ideal tools to study bone sarcomas.

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Comparative proteome analysis of monolayer and spheroid culture of canine osteosarcoma cells

Cited by 16 publications

References 70 publications

Proteomic profiling and identification of significant markers from high-grade osteosarcoma after cryotherapy and irradiation

Proteomic profiling and identification of significant markers from high-grade osteosarcoma after cryotherapy and irradiation

The Differences in the Proteome Profile of Cannabidiol-Treated Skin Fibroblasts following UVA or UVB Irradiation in 2D and 3D Cell Cultures

In vitro three-dimensional cell cultures for bone sarcomas

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