Characterization of Changes in the Proteome in Different Regions of 3D Multicell Tumor Spheroids

被引：58

作者：

McMahon, K. M. ^{[1
]}

Volpato, M. ^{[1
]}

Chi, H. Y. ^{[1
]}

Musiwaro, P. ^{[1
]}

Poterlowicz, K. ^{[2
]}

Peng, Y. ^{[2
]}

Scally, A. J. ^{[3
]}

Patterson, L. H. ^{[1
]}

Phillips, R. M. ^{[1
]}

Sutton, C. W. ^{[1
]}

机构：

[1] Univ Bradford, Inst Canc Therapeut, Bradford BD7 1DP, W Yorkshire, England

[2] Univ Bradford, Dept Comp, Bradford BD7 1DP, W Yorkshire, England

[3] Univ Bradford, Sch Hlth Studies, Bradford BD7 1DP, W Yorkshire, England

来源：

JOURNAL OF PROTEOME RESEARCH | 2012年 / 11卷 / 05期

基金：

英国工程与自然科学研究理事会;

关键词：

cancer; spheroids; proteomics; autophagy; necrosis; hypoxia; metabolic processes; lipid metabolism; CYCLE INHIBITOR P27(KIP1); BIOREDUCTIVE DRUGS; CELL-CULTURE; EXPRESSION; HYPOXIA; MONOLAYER; CARCINOMA; NECROSIS; GLUCOSE; MODELS;

D O I：

10.1021/pr2012472

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Three dimensional multicell tumor spheroids (MCTS) provide an experimental model where the influence of microenvironmental conditions on protein expression can be determined. Sequential trypsin digestion of HT29 colon carcinoma MCTS enabled segregation into four populations comprising proliferating cells from the surface (SL), an intermediate region (IR), nonproliferating hypoxic cells from the perinecrotic region (PN), and a necrotic core (NC). Total protein was extracted from each population and subjected to iTRAQ-based quantitative proteomics analysis. From a total of 887 proteins identified, 209 were observed to be up-regulated and 114 were down-regulated in the PN and NC regions relative to the SL. Among the up-regulated proteins, components of glycolysis, TCA cycle, lipid metabolism, and steroid biosynthesis increased progressively toward the PN and NC regions. Western blotting, immunohistochemistry, and enzyme assays confirmed that significant changes in the expression of proteins involved in cellular metabolism occur in the nonproliferating fraction of cells within the viable rim. The presence of full length, functional proteins within the NC was unexpected, and further analysis demonstrated that this region contains cells that are undergoing autophagy. This study has identified possible targets that may be suitable for therapeutic intervention, and further studies to validate these are required.

引用

页码：2863 / 2875

页数：13

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