Dynamics of signaling, cytoskeleton and cell cycle regulation proteins in glioblastoma cells after sub-lethal photodynamic treatment: Antibody microarray study

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• 作者：Anatoly Uzdensky^a ; ^{auzd@yandex.ru} ; Bjorn Kristiansen^b ; Johan Moan^c ; ^d ; Asta Juzeniene^c
• 关键词：Proteomics ; Antibody microarray ; Signaling ; Cytoskeleton ; Proliferation ; Adhesion
• 刊名：Biochimica et Biophysica Acta (BBA)/General Subjects
• 出版年：2012
• 出版时间：July, 2012
• 年：2012
• 卷：1820
• 期：7
• 页码：795-803
• 全文大小：612 K

文摘

Background

Photodynamic therapy (PDT) that induces oxidative stress and cell death is used for tumor destruction in oncology. To characterize early molecular events in photosensitized glioblastoma cells, we studied expression of 224 proteins after sublethal PDT that doesn't kill but wounds cells.

Methods

Cultured glioblastoma D54Mg cells were photosensitized with 5-aminolevulinic acid so that cell survival was 95-100 % . At following 0.5-5.5 h protein expression and phosphorylation was assayed using proteomic antibody microarrays.

Results

Within the first post-treatment hour we observed phosphorylation of protein kinase Raf, adhesion-related kinases FAK and Pyk2, and microtubule-associated protein tau. Protein kinase C¦Ã and microtubule-associated protein MAP-1B were overexpressed. Dystrophin, calponin, and vinculin, components of the actin cytoskeleton scaffold, microtubule-associated proteins MAP2 and CNP, cytokeratins 4 and 7 were down-regulated that indicated changes in adhesion and cell shape. Down-regulation of cyclins A, D1 and D3, c-Myc, checkpoint proteins chk1/2 and up-regulation of Smad4 could arrest the cell cycle. Overexpression of Bcl-xL and down-regulation of caspase 9 demonstrated anti-apoptotic response. At 2 h post-treatment protein expression changed lesser but at 5.5 h levels of PKC¦Ã and ¦Â-synuclein and phosphorylation of Raf, FAK, Pyk2, and tau increased again.

Conclusions

Sub-lethal PDT induces complex response of glioblastoma cells including changes in activity and expression of proteins involved in adhesion-mediated signaling, signal transduction, cytoskeleton remodeling, cell cycle regulation and anti-apoptotic processes.

General significance

Multiple reactions of various cellular subsystems including adhesion, cytoskeleton, signal transduction, cell cycle, and apoptosis are integrated into the general cell response to a sublethal impact.