1. Identification of Up- and Down-Regulated Proteins in Pemetrexed-Resistant Human Lung Adenocarcinoma: Flavin Reductase and Calreticulin Play Key Roles in the Development of Pemetrexed-Associated Resistance
- Author
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Ying-Jen Chen, Hsiu-Chuan Chou, Ting-Wen Chung, En-Chi Liao, Yu-Shan Wei, Hong-Lin Chan, Yi-Ting Tsai, Dai-Ying Lin, Ching-Hsiung Lin, Yueh-Feng Wen, Chi-Chen Lin, Sheng-Hao Lin, and Jing-Yi Chen
- Subjects
Proteomics ,Lung Neoplasms ,FMN Reductase ,Proteome ,Cell Survival ,Adenocarcinoma of Lung ,Antineoplastic Agents ,Apoptosis ,Pemetrexed ,Biology ,Adenocarcinoma ,Biochemistry ,Cell Line, Tumor ,Flavin reductase ,medicine ,Humans ,Electrophoresis, Gel, Two-Dimensional ,Viability assay ,RNA, Small Interfering ,A549 cell ,General Chemistry ,Molecular biology ,Neoplasm Proteins ,Gene Expression Regulation, Neoplastic ,Drug Resistance, Neoplasm ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,biology.protein ,Signal transduction ,Calreticulin ,medicine.drug ,Signal Transduction - Abstract
Drug resistance is one of the major causes of cancer chemotherapy failure. In the current study, we used a pair of lung adenocarcinoma cell lines, A549 and the pemetrexed-resistant A549/PEM cells, as a model to monitor resistance-dependent cellular responses and identify potential therapeutic targets. By means of 2D differential gel electrophoresis (2D-DIGE) and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), we investigated the global protein expression alterations induced by pemetrexed treatment and resistance. The proteomic result revealed that pemetrexed exposure obviously altered the expression of 81 proteins in the A549 cells, whereas no significant response was observed in the similarly treated A549/PEM cells, hence implying an association between these proteins and the drug-specific response. Moreover, 72 proteins including flavin reductase and calreticulin demonstrated differential expression between the A549 and A549/PEM cells, indicating baseline resistance. Additional tests employed siRNA silencing, protein overexpression, cell viability analysis, and analysis of apoptosis to examine and confirm the potency of flavin reductase and calreticulin proteins in the development of pemetrexed resistance. In summary, by using a proteomic approach, we identified numerous proteins, including flavin reductase and calreticulin, involved in pemetrexed drug resistance-developing mechanisms. Our results provide useful diagnostic markers and therapeutic candidates for pemetrexed-resistant lung cancer treatment.
- Published
- 2015