Proteomics

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Secretome analysis of glioblastoma cells

ABSTRACT: PKR-like kinase (PERK) plays a significant role in inducing angiogenesis in various cancer-types including glioblastoma. Mass spectrometry screen on conditioned media from LN308 glioblastoma cell line treated with hypoxia revealed Peptidylglycine α-amidating monooxygenase (PAM) as a potential target inducing angiogenesis. PERK is found to be regulating PAM at mRNA level. PERK activation via CCT020312 (PERK activator) also increased the cleavage and thus the generation of sfCD domain of PAM in an oxygen-independent manner which acts as a signaling molecule from cytoplasm to the nuclei. PERK was found interacting with PAM in immuno-precipitation experiments suggesting a possible involvement in the generation of the PAM sfCD domain. Knockdown of PERK or PAM in LN308 cells reduced the formation of tubes by HUVECs in vitro. In vivo data also highlighted the importance of PAM in growth of glioblastoma where reduction of PAM expression in engrafted tumor significantly increased the survival in mice. In summary, PAM acts as a potential target for therapy against angiogenesis in glioblastoma.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Chi Nguyen  

LAB HEAD: Robert Ahrends

PROVIDER: PXD012523 | Pride | 2020-03-05

REPOSITORIES: Pride

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20160329_Secretome_GSK_Hypoxia_phenyx_1.mgf Mgf
20160329_Secretome_GSK_Hypoxia_phenyx_1.pride.mgf.gz Mgf
20160329_Secretome_GSK_Hypoxia_phenyx_10.mgf Mgf
20160329_Secretome_GSK_Hypoxia_phenyx_10.pride.mgf.gz Mgf
20160329_Secretome_GSK_Hypoxia_phenyx_11.mgf Mgf
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Publications

PERK-mediated expression of peptidylglycine α-amidating monooxygenase supports angiogenesis in glioblastoma.

Soni Himanshu H   Bode Julia J   Nguyen Chi D L CDL   Puccio Laura L   Neßling Michelle M   Piro Rosario M RM   Bub Jonas J   Phillips Emma E   Ahrends Robert R   Eipper Betty A BA   Tews Björn B   Goidts Violaine V  

Oncogenesis 20200213 2

PKR-like kinase (PERK) plays a significant role in inducing angiogenesis in various cancer types including glioblastoma. By proteomics analysis of the conditioned medium from a glioblastoma cell line treated with a PERK inhibitor, we showed that peptidylglycine α-amidating monooxygenase (PAM) expression is regulated by PERK under hypoxic conditions. Moreover, PERK activation via CCT020312 (a PERK selective activator) increased the cleavage and thus the generation of PAM cleaved cytosolic domain  ...[more]

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