Nanoproteomic analysis of ischemia-dependent changes in signaling protein phosphorylation in colorectal normal and cancer tissue

Journal of Translational Medicine

Research

Carolyn Compton¹  Helen Moore²  Lokesh Agrawal²  Florian T. Unger³  Kerstin A. David³  Nicole Lange³  Jana Krüger³  Hartmut Juhl³ 

[1] Arizona State University, Phoenix, AZ, USA;Biorepositories and Biospecimen Research Branch,National Cancer Institute, National Institutes of Health, Bethesda, MD, USA;Indivumed GmbH, Falkenried 88, 20251, Hamburg, Germany;

关键词: Clinical biospecimen;    Ischemia;    NanoPro1000;    Isoform phosphorylation;    Clinical testing;   

DOI  :  10.1186/s12967-015-0752-1

 received in 2015-07-17, accepted in 2015-12-02,  发布年份 2016

来源: Springer

【 摘 要 】

BackgroundClinical diagnostic research relies upon the collection of tissue samples, and for those samples to be representative of the in vivo situation. Tissue collection procedures, including post-operative ischemia, can impact the molecular profile of the tissue at the genetic and proteomic level. Understanding the influence of factors such as ischemia on tissue samples is imperative in order to develop both markers of tissue quality and ultimately accurate diagnostic tests.MethodsUsing NanoPro1000 technology, a rapid and highly sensitive immunoassay platform, the phosphorylation status of clinically relevant cancer-related biomarkers in response to ischemia was quantified in tissue samples from 20 patients with primary colorectal cancer. Tumor tissue and adjacent normal tissue samples were collected and subjected to cold ischemia prior to nanoproteomic analysis of AKT, ERK1/2, MEK1/2, and c-MET. Ischemia-induced relative changes in overall phosphorylation and phosphorylation of individual isoforms were calculated and statistical significance determined. Any differences in baseline levels of phosphorylation between tumor tissue and normal tissue were also analyzed.ResultsChanges in overall phosphorylation of the selected proteins in response to ischemia revealed minor variations in both normal and tumor tissue; however, significant changes were identified in the phosphorylation of individual isoforms. In normal tissue post-operative ischemia, phosphorylation was increased in two AKT isoforms, two ERK1/2 isoforms, and one MEK1/2 isoform and decreased in one MEK1/2 isoform and one c-MET isoform. Following ischemia in tumor tissue, one AKT isoform showed decreased phosphorylation and there was an overall increase in unphosphorylated ERK1/2, whereas an increase in the phosphorylation of two MEK1/2 isoforms was observed. There were no changes in c-MET phosphorylation in tumor tissue.ConclusionThis study provides insight into the influence of post-operative ischemia on tissue sample biology, which may inform the future development of markers of tissue quality and assist in the development of diagnostic tests.

【 授权许可】

CC BY   
© Unger et al. 2016