Accumulation of an anthraquinone and its platinum complexes in cancer cell spheroids: the effect of charge on drug distribution in solid tumour models

Cell cultures are widely used model systems. Some immortalized cell lines can be grown in either two-dimensional (2D) adherent monolayers or in three-dimensional (3D) multicellular aggregates, or spheroids. Here, the quantitative proteome and phosphoproteome of colon carcinoma HT29 cells cultures in 2D monolayers and 3D spheroids were compared with a stable isotope labeling of amino acids (SILAC) labeling strategy. Two biological replicates from each sample were examined and notable differences in both the proteome and the phosphoproteome were determined by nano-liquid chromatography tandem mass spectrometry (nLC-MS/MS) to assess how growth configuration affects molecular expression. A total of 5867 protein groups, including 2523 phosphoprotein groups and 8733 phosphopeptides were identified in the samples. The Gene Ontology analysis revealed enriched GO terms in the 3D samples for RNA binding, nucleic acid binding, enzyme binding, cytoskeletal protein binding, and histone binding for their molecular functions (MF) and in the process of cell cycle, cytoskeleton organization, and DNA metabolic process for the biological process (BP). The KEGG pathway analysis indicated that 3D cultures are enriched for oxidative phosphorylation pathways, metabolic pathways, peroxisome pathways, and biosynthesis of amino acids. In contrast, analysis of the phosphoproteomes indicated that 3D cultures have decreased phosphorylation correlating with slower growth rates and lower cell-to extracellular matrix interactions. In sum, these results provide quantitative assessments of the effects on the proteome and phosphoproteome of culturing cells in 2D versus 3D cell culture configurations.

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“…(14) 3D cell cultures are also widely used to evaluate the penetration and metabolism of new drug compounds. (10, 15–18)…”

Section: Introductionmentioning

confidence: 99%

Quantitative Proteomic and Phosphoproteomic Comparison of 2D and 3D Colon Cancer Cell Culture Models

et al. 2016

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“…However these systems are often studied using two dimensional (2D) cell monolayers which cannot reproduce the complex three dimensional (3D) environment in tissues or organs. To better model the actual in vivo conditions, 3D multicellular spheroids were developed and they have been used as avascular tumour models for evaluating small molecule [16] or nanoparticle [17,18] delivery. Magnetic multicellular spheroids could be generated from magnetically labelled cells.…”

Section: Introductionmentioning

confidence: 99%

pH-responsive endosomolytic pseudo-peptides for drug delivery to multicellular spheroids tumour models

Slater

Chen

2011

Biomaterials

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Endosomolytic polymers can aid in the endosomal release of therapeutics to improve intracellular drug delivery. pH-responsive biomimetic pseudo-peptides were synthesised by grafting L-phenylalanine onto the pendant carboxylic acids of a polyamide, poly(L-lysine isophthalamide). PP-75 (stoichiometric L-phenylalanine grafting of 75 mol%) was determined to have the best endosomolytic property. The mean hydrodynamic size of PP-75 decreased with lower pH as the polymers adopted a more compact conformation due to protonation of acidic groups and increase in hydrophobicity. PP-75 was demonstrated to deliver model drugs effectively in three dimensional (3D) magnetic HeLa multicellular spheroids used as in vitro tumour models. These spheroids can be isolated easily and quickly by magnetic separation.Due to its relatively small size, PP-75 was able to penetrate from the exterior to the interior of these spheroids and was internalised by the cells in the spheroids. It could retain its pH-mediated membrane-lytic capability in 3D drug delivery by releasing internalised calcein from intracellular endosomes in the tumour models. Furthermore, cell viability results suggest that PP-75 showed no significant cytotoxicity towards cells in the spheroids. The pH-responsive PP-75 can potentially enhance the extracellular and intracellular delivery of therapeutics in tumours.

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“…Factors causing MCR include: intercellular molecular changes like activation of Stat3 and NF-κB signaling pathways, microenvironment alterations like changes in extracellular matrix (ECM) and varying pH, the presence of hypoxic cells and differences in cell cycle distribution within MCTS, and importantly, drug penetration within spheroids, which is the rate-limiting step for drug delivery 104,111–112 . In addition to structural and physical properties, poor penetration of drug molecules into spheroids can be caused by binding with the outer layers of cells or physical barriers due to the ECM and cellular membranes 108,113–114 . Additionally, pH gradients within the 3D structure sometimes protect cells in the acidic core from weakly acidic compounds such as mitoxantrones, while enhancing the effect of weakly basic drugs such as chlorambucil 115 .…”

Section: A Novel Application: 3d Cell Culture Systemsmentioning

confidence: 99%

Mass Spectrometry Imaging of Therapeutics from Animal Models to Three-Dimensional Cell Cultures

Liu

Hummon

2015

Anal. Chem.

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Mass spectrometry imaging (MSI) is a powerful label-free technique for the investigation of the spatial distribution of molecules at complex surfaces and has been widely used in the pharmaceutical sciences to understand the distribution of different drugs and their metabolites in various biological samples, ranging from cell-based models to tissues. Here, we review the current applications of MSI for drug studies in animal models, followed by a discussion of the novel advances of MSI in three-dimensional (3D) cell cultures for accurate, efficient and high-throughput analyses to evaluate therapeutics.

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Accumulation of an anthraquinone and its platinum complexes in cancer cell spheroids: the effect of charge on drug distribution in solid tumour models

Cited by 70 publications

References 13 publications

Quantitative Proteomic and Phosphoproteomic Comparison of 2D and 3D Colon Cancer Cell Culture Models

Quantitative Proteomic and Phosphoproteomic Comparison of 2D and 3D Colon Cancer Cell Culture Models

pH-responsive endosomolytic pseudo-peptides for drug delivery to multicellular spheroids tumour models

Mass Spectrometry Imaging of Therapeutics from Animal Models to Three-Dimensional Cell Cultures

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