Proteome and Acetylome Analysis Identifies Novel Pathways and Targets Regulated by Perifosine in Neuroblastoma

Xiao Gu,Wei Tian,Zhihui Liu,Yudi Dong,Yue Zhan,Xiaowen Zhang,Zhijie Li,Zhongyan Hua,Carol J Thiele

doi:10.1038/srep42062

Abstract

Perifosine, an Akt inhibitor, has been shown to be effective in controlling neuroblastoma tumor growth. However, studies indicate that in addition to the ability to inhibit Akt, other mechanisms contribute to perifosine’s anti-tumor activity. To gain insight into perifosine anti-tumor activity in neuroblastoma we have studied changes in the proteome and acetylome after perifosine treatment in SK-N-AS neuroblastoma cells using SILAC labeling, affinity enrichment, high-resolution and LC-MS/MS analysis. Bioinformatic analysis indicates that, a total of 5,880 proteins and 3,415 lysine acetylation sites were quantified in SK-N-AS cells and 216 differentially expressed proteins and 115 differentially expressed lysine acetylation sites were obtained. These differentially expressed proteins and lysine acetylated proteins were involved in a number of different biological functions, metabolic pathways and pathophysiological processes. This study details the impact of perifosine on proteome and lysine acetylome in SK-N-AS cells and expands our understanding of the mechanisms of perifosine action in neuroblastoma.

Highlights

Akt activation is detected in the tumor tissues of patients with high-risk NB, and is a predictor of poor prognosis[7]
We explored the mechanisms underlying perifosine treatment and our results provided novel insights into the action and acetylation caused by perifosine
We treated AS cells with varying concentrations (2.5, 5, 7.5, 10 and μM) of perifosine for h to study the effect of perifosine on the expressions of phosphorylated-Akt and protein acetylation

Summary

Introduction

Akt activation is detected in the tumor tissues of patients with high-risk NB, and is a predictor of poor prognosis[7]. Such studies indicate that Akt may be a potential therapeutic target for NB. To explore the mechanisms underlying perifosine treatment, the global proteome and lysine acetylome profile in SK-N-AS (AS) cells after perifosine treatment were intensively studied by a combination of www.nature.com/scientificreports/. We obtained 216 differentially expressed proteins from the 5,880 quantified proteins, and 115 differentially expressed lysine acetylation sites from the 3,415 quantified lysine acetylation sites These proteins mainly exist in the nucleus and cytoplasm, and are involved in a wide variety of metabolic pathways and biological functions, such as lipid metabolic process, apoptosis and transcription. We explored the mechanisms underlying perifosine treatment and our results provided novel insights into the action and acetylation caused by perifosine

Methods

Results

Conclusion