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Reverse-Phase Protein Array Analysis to Identify Biomarker Proteins in Human Pancreatic Cancer

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Abstract

Background

Pancreatic cancer is the fourth leading cause of cancer death in the United States. The high mortality rate of patients with pancreatic cancer is primarily due to the difficulty of early diagnosis and a lack of effective therapies. There is an urgent need to discover novel molecular targets for early diagnosis and new therapeutic approaches to improve the clinical outcome of this deadly disease.

Aim

We utilized the reverse-phase protein assay (RPPA) to identify differentially expressed biomarker proteins in tumors and matched adjacent, normal-appearing tissue samples from 15 pancreatic cancer patients.

Methods

The antibody panel used for the RPPA included 130 key proteins involved in various cancer-related pathways. The paired t test was used to determine the significant differences between matched pairs, and the false discovery rate-adjusted p values were calculated to take into account the effect of multiple comparisons.

Results

After correcting for multiple comparisons, we found 19 proteins that had statistically significant differences in expression between matched pairs. However, only four (AKT, β-catenin, GAB2, and PAI-1) of them met the conservative criteria (both a q value <0.05 and a fold-change of ≥3/2 or ≤2/3) to be considered differentially expressed. Overexpression of AKT, β-catenin, and GAB2 in pancreatic cancer tissues identified by RPPA has also been further confirmed by western blot analysis. Further analysis identified several significantly associated canonical pathways and overrepresented network functions.

Conclusion

GAB2, a newly identified protein in pancreatic cancer, may provide additional insight into this cancer’s pathogenesis. Future studies in a larger population are warranted to further confirm our results.

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Acknowledgments

This work was supported by the National Institutes of Health through the University of Texas MD Anderson Cancer Center Support Grant CA016672 (R. DePinho), research grant R03 CA132103 (C. Wei), and Seed Funding Research Program Grant from Duncan Family Institute in the University of Texas MD Anderson Cancer Center (C. Wei). We thank Diane Hackett and Jill Delsigne for their editorial comments.

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Authors and Affiliations

  1. Department of Epidemiology, Unit 1365, The University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, TX, 77030, USA

    Yu-Jing Huang, Marsha L. Frazier, Qian Liu & Chongjuan Wei

  2. Department of Bioinformatics and Computational Biology, The University of Texas MD Anderson Cancer Center, Houston, TX, 77030, USA

    Nianxiang Zhang

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  1. Yu-Jing Huang
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Correspondence to Chongjuan Wei.

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Huang, YJ., Frazier, M.L., Zhang, N. et al. Reverse-Phase Protein Array Analysis to Identify Biomarker Proteins in Human Pancreatic Cancer. Dig Dis Sci 59, 968–975 (2014). https://doi.org/10.1007/s10620-013-2938-9

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  • DOI: https://doi.org/10.1007/s10620-013-2938-9

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