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(Chest. 2002;121:28S.)
© 2002 American College of Chest Physicians

Proteomic Analysis of Mouse Lung Neoplasia*

Katherine Peebles, BS; Kim Fung, BS; Al Malkinson, PhD and Mark Duncan, PhD

* From the Department of Pharmaceutical Sciences, University of Colorado Health Sciences Center, Denver, CO.

Correspondence to: Katherine Peebles, BS, Department of Pharmaceutical Sciences, School of Pharmacy, University of Colorado Health Sciences Center, Campus Box C238, 4200 E 9th Ave, Denver, CO 80262

Proteomics has the potential to significantly advance the understanding of biological systems. In this study, we applied classic and more novel proteomics techniques to in vitro and in vivo models of lung cancer. The traditional comparison of protein expression entails the separation of proteins by two-dimensional gel electrophoresis (2DE) and subsequent mass fingerprinting by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOFMS). Separations based on 2DE revealed at least 20 spots that differed between a nontumorigenic mouse lung epithelial cell line (E10) and its spontaneous transformant (E9). Differentially expressed proteins identified by this technique include lipocortin I, which is of great interest because of the role of inflammation in lung tumorigenesis. Since 2DE is a time-consuming process and detects only a small fraction of the proteins expressed in cells or tissue, we also employed a combination of MALDI-TOFMS of whole-tissue homogenates and electrospray ionization liquid chromatography coupled to tandem mass spectrometry. This was applied to total pools of trypsinized proteins to compare protein expression in urethane-induced mouse lung adenomas with healthy lung tissue. Thus far, we have identified > 400 proteins from both the 10,000g fraction and the nuclear fraction of the tumor sample, and > 200 proteins from healthy tissue. By utilizing both of these techniques, we have attempted to provide a thorough description of the changes in protein expression that occur during lung carcinogenesis.


   Footnotes
 
Abbreviations: 2DE = two-dimensional gel electrophoresis; MALDI-TOFMS = matrix-assisted laser desorption ionization time of flight mass spectrometry

This research was supported by US Public Health Service grant CA33497.





This Article
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Right arrow Articles by Duncan, M.

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