Identification of differentially expressed genes in experimental models of the tumor microenvironment using differential display

H J Knowles, R M Phillips

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

BACKGROUND: Genes upregulated within the tumour microenvironment represent potential targets for rational drug design. Most studies to date concentrate on the effects of hypoxia, although it is likely many genes are regulated by a more physiological combination of factors.

MATERIALS & METHODS: Cells under conditions analogous to the normal and tumour microenvironments were isolated from the plateau-phase system and multicellular spheroids. Gene expression was analysed by differential display and confirmed by Northern blot or semiquantitative RT-PCR.

RESULTS: p21-activated kinase (PAK1), a calmodulin-related mRNA, cytochrome oxidase subunit I and an H3.3 histone were upregulated within the in vitro tumour microenvironment, the last 3 within spheroids.

CONCLUSIONS: Both models exhibit a range of microenvironmental parameters, although spheroids are more physiological with respect to the presence of extreme hypoxia and the formation of 3-dimensional interactions. We have shown that it is feasible to manipulate the spheroid system by serial trypsinisation to obtain reproducible cell populations for gene expression studies.

LanguageEnglish
Pages2305-2311
Number of pages7
JournalAnticancer Research
Volume21
Issue number4A
Publication statusPublished - 29 Nov 2001
Externally publishedYes

Fingerprint

Tumor Microenvironment
Theoretical Models
Cellular Spheroids
p21-Activated Kinases
Genes
Gene Expression
Drug Design
Electron Transport Complex IV
Calmodulin
Northern Blotting
Histones
Polymerase Chain Reaction
Messenger RNA
Population
Hypoxia

Cite this

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title = "Identification of differentially expressed genes in experimental models of the tumor microenvironment using differential display",
abstract = "BACKGROUND: Genes upregulated within the tumour microenvironment represent potential targets for rational drug design. Most studies to date concentrate on the effects of hypoxia, although it is likely many genes are regulated by a more physiological combination of factors.MATERIALS & METHODS: Cells under conditions analogous to the normal and tumour microenvironments were isolated from the plateau-phase system and multicellular spheroids. Gene expression was analysed by differential display and confirmed by Northern blot or semiquantitative RT-PCR.RESULTS: p21-activated kinase (PAK1), a calmodulin-related mRNA, cytochrome oxidase subunit I and an H3.3 histone were upregulated within the in vitro tumour microenvironment, the last 3 within spheroids.CONCLUSIONS: Both models exhibit a range of microenvironmental parameters, although spheroids are more physiological with respect to the presence of extreme hypoxia and the formation of 3-dimensional interactions. We have shown that it is feasible to manipulate the spheroid system by serial trypsinisation to obtain reproducible cell populations for gene expression studies.",
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Identification of differentially expressed genes in experimental models of the tumor microenvironment using differential display. / Knowles, H J; Phillips, R M.

In: Anticancer Research, Vol. 21, No. 4A, 29.11.2001, p. 2305-2311.

Research output: Contribution to journalArticle

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AU - Phillips, R M

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AB - BACKGROUND: Genes upregulated within the tumour microenvironment represent potential targets for rational drug design. Most studies to date concentrate on the effects of hypoxia, although it is likely many genes are regulated by a more physiological combination of factors.MATERIALS & METHODS: Cells under conditions analogous to the normal and tumour microenvironments were isolated from the plateau-phase system and multicellular spheroids. Gene expression was analysed by differential display and confirmed by Northern blot or semiquantitative RT-PCR.RESULTS: p21-activated kinase (PAK1), a calmodulin-related mRNA, cytochrome oxidase subunit I and an H3.3 histone were upregulated within the in vitro tumour microenvironment, the last 3 within spheroids.CONCLUSIONS: Both models exhibit a range of microenvironmental parameters, although spheroids are more physiological with respect to the presence of extreme hypoxia and the formation of 3-dimensional interactions. We have shown that it is feasible to manipulate the spheroid system by serial trypsinisation to obtain reproducible cell populations for gene expression studies.

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KW - Cell Division/physiology

KW - Cell Hypoxia/genetics

KW - Colonic Neoplasms/genetics

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KW - Gene Expression Regulation, Neoplastic

KW - Humans

KW - Reverse Transcriptase Polymerase Chain Reaction

KW - Spheroids, Cellular

KW - Trypsin/pharmacology

KW - Tumor Cells, Cultured

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