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Molecular and phenotypic characterisation of paediatric glioma cell lines as models for preclinical drug development.

Bax DA, Little SE, Gaspar N, Perryman L, Marshall L, Viana-Pereira M, Jones TA, Williams RD, Grigoriadis A, Vassal G, Workman P, Sheer D, Reis RM, Pearson AD, Hargrave D, Jones C - PLoS ONE (2009)

Bottom Line: All lines proliferate as adherent monolayers and express glial markers.Copy number profiling revealed complex genomes including amplification and deletions of genes known to be pivotal in core glioblastoma signalling pathways.Expression profiling identified 93 differentially expressed genes which were able to distinguish between the adult and paediatric high grade cell lines, including a number of kinases and co-ordinated sets of genes associated with DNA integrity and the immune response.

View Article: PubMed Central - PubMed

Affiliation: Paediatric Oncology, The Institute of Cancer Research, Sutton, United Kingdom.

ABSTRACT

Background: Although paediatric high grade gliomas resemble their adult counterparts in many ways, there appear to be distinct clinical and biological differences. One important factor hampering the development of new targeted therapies is the relative lack of cell lines derived from childhood glioma patients, as it is unclear whether the well-established adult lines commonly used are representative of the underlying molecular genetics of childhood tumours. We have carried out a detailed molecular and phenotypic characterisation of a series of paediatric high grade glioma cell lines in comparison to routinely used adult lines.

Principal findings: All lines proliferate as adherent monolayers and express glial markers. Copy number profiling revealed complex genomes including amplification and deletions of genes known to be pivotal in core glioblastoma signalling pathways. Expression profiling identified 93 differentially expressed genes which were able to distinguish between the adult and paediatric high grade cell lines, including a number of kinases and co-ordinated sets of genes associated with DNA integrity and the immune response.

Significance: These data demonstrate that glioma cell lines derived from paediatric patients show key molecular differences to those from adults, some of which are well known, whilst others may provide novel targets for evaluation in primary tumours. We thus provide the rationale and demonstrate the practicability of using paediatric glioma cell lines for preclinical and mechanistic studies.

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Related in: MedlinePlus

Constitutive activation of key signalling pathways in glioma cell lines.Western blots analysis of c-Raf, phospho/total Erk1/2, phospho/total Akt, phospho/total GSK3β, phospho/total S6 and GAPDH as loading control in adult (LN229, A172, U118MG, U138MG, U87MG, SF268) and paediatric (SF188, KNS42, UW479, Res259, Res186) glioma cell lines.
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pone-0005209-g003: Constitutive activation of key signalling pathways in glioma cell lines.Western blots analysis of c-Raf, phospho/total Erk1/2, phospho/total Akt, phospho/total GSK3β, phospho/total S6 and GAPDH as loading control in adult (LN229, A172, U118MG, U138MG, U87MG, SF268) and paediatric (SF188, KNS42, UW479, Res259, Res186) glioma cell lines.

Mentions: Protein expression was measured by Western blot to examine constitutive pathway activation of several key signalling transduction networks of importance in gliomagenesis (Figure 3). There was a modest activation of the MAPK pathway as determined by levels of phosphorylated Erk 1/2 in the paediatric high grade lines SF188 and UW479 as compared to KNS42 and the low grade lines (Res259, Res186). Although Akt pathway activation was seen in all paediatric cells, this was considerably lower than that observed in our panel of adult glioblastoma lines, except in the PTEN line Res186, which showed high levels of phospho-Akt. Phosphorylated GSK3β levels were low in UW479, Res259 and Res186, and almost entirely absent in the glioblastoma lines SF188 and KNS42. Although there was some degree of ribosomal protein S6 activation in all lines, this was particularly pronounced in KNS42 and Res259, with very high levels of phospho-S6. Taken together, although variable, we observed constitutive activation of one or more of these signal transduction pathways in all the paediatric glioma lines.


Molecular and phenotypic characterisation of paediatric glioma cell lines as models for preclinical drug development.

Bax DA, Little SE, Gaspar N, Perryman L, Marshall L, Viana-Pereira M, Jones TA, Williams RD, Grigoriadis A, Vassal G, Workman P, Sheer D, Reis RM, Pearson AD, Hargrave D, Jones C - PLoS ONE (2009)

Constitutive activation of key signalling pathways in glioma cell lines.Western blots analysis of c-Raf, phospho/total Erk1/2, phospho/total Akt, phospho/total GSK3β, phospho/total S6 and GAPDH as loading control in adult (LN229, A172, U118MG, U138MG, U87MG, SF268) and paediatric (SF188, KNS42, UW479, Res259, Res186) glioma cell lines.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2666263&req=5

pone-0005209-g003: Constitutive activation of key signalling pathways in glioma cell lines.Western blots analysis of c-Raf, phospho/total Erk1/2, phospho/total Akt, phospho/total GSK3β, phospho/total S6 and GAPDH as loading control in adult (LN229, A172, U118MG, U138MG, U87MG, SF268) and paediatric (SF188, KNS42, UW479, Res259, Res186) glioma cell lines.
Mentions: Protein expression was measured by Western blot to examine constitutive pathway activation of several key signalling transduction networks of importance in gliomagenesis (Figure 3). There was a modest activation of the MAPK pathway as determined by levels of phosphorylated Erk 1/2 in the paediatric high grade lines SF188 and UW479 as compared to KNS42 and the low grade lines (Res259, Res186). Although Akt pathway activation was seen in all paediatric cells, this was considerably lower than that observed in our panel of adult glioblastoma lines, except in the PTEN line Res186, which showed high levels of phospho-Akt. Phosphorylated GSK3β levels were low in UW479, Res259 and Res186, and almost entirely absent in the glioblastoma lines SF188 and KNS42. Although there was some degree of ribosomal protein S6 activation in all lines, this was particularly pronounced in KNS42 and Res259, with very high levels of phospho-S6. Taken together, although variable, we observed constitutive activation of one or more of these signal transduction pathways in all the paediatric glioma lines.

Bottom Line: All lines proliferate as adherent monolayers and express glial markers.Copy number profiling revealed complex genomes including amplification and deletions of genes known to be pivotal in core glioblastoma signalling pathways.Expression profiling identified 93 differentially expressed genes which were able to distinguish between the adult and paediatric high grade cell lines, including a number of kinases and co-ordinated sets of genes associated with DNA integrity and the immune response.

View Article: PubMed Central - PubMed

Affiliation: Paediatric Oncology, The Institute of Cancer Research, Sutton, United Kingdom.

ABSTRACT

Background: Although paediatric high grade gliomas resemble their adult counterparts in many ways, there appear to be distinct clinical and biological differences. One important factor hampering the development of new targeted therapies is the relative lack of cell lines derived from childhood glioma patients, as it is unclear whether the well-established adult lines commonly used are representative of the underlying molecular genetics of childhood tumours. We have carried out a detailed molecular and phenotypic characterisation of a series of paediatric high grade glioma cell lines in comparison to routinely used adult lines.

Principal findings: All lines proliferate as adherent monolayers and express glial markers. Copy number profiling revealed complex genomes including amplification and deletions of genes known to be pivotal in core glioblastoma signalling pathways. Expression profiling identified 93 differentially expressed genes which were able to distinguish between the adult and paediatric high grade cell lines, including a number of kinases and co-ordinated sets of genes associated with DNA integrity and the immune response.

Significance: These data demonstrate that glioma cell lines derived from paediatric patients show key molecular differences to those from adults, some of which are well known, whilst others may provide novel targets for evaluation in primary tumours. We thus provide the rationale and demonstrate the practicability of using paediatric glioma cell lines for preclinical and mechanistic studies.

Show MeSH
Related in: MedlinePlus