title: 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.
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.
authors.affiliation: Department of Applied Science, National Hsinchu University of Education , No. 521, Nan-Da Road, Hsinchu 30014, Taiwan.
authors.forename: Hsiu-Chuan
authors.initials: HC
authors.lastname: Chou
authors.affiliation: Department of Medical Science and Institute of Bioinformatics and Structural Biology, National Tsing Hua University , No. 101, Kuang-Fu Road Sec. 2, Hsinchu 30013, Taiwan.
authors.forename: Jing-Yi
authors.initials: JY
authors.lastname: Chen
authors.affiliation: Department of Medical Science and Institute of Bioinformatics and Structural Biology, National Tsing Hua University , No. 101, Kuang-Fu Road Sec. 2, Hsinchu 30013, Taiwan.
authors.forename: Dai-Ying
authors.initials: DY
authors.lastname: Lin
authors.affiliation: Department of Medical Science and Institute of Bioinformatics and Structural Biology, National Tsing Hua University , No. 101, Kuang-Fu Road Sec. 2, Hsinchu 30013, Taiwan.|Department of Internal Medicine, National Taiwan University Hospital Hsinchu Branch , No. 25, Lane 442, Sec. 1, Jingguo Road, Hsinchu 30059, Taiwan.
authors.forename: Yueh-Feng
authors.initials: YF
authors.lastname: Wen
authors.affiliation: Institute of Biomedical Science, National Chung-Hsing University , No. 250, Guoguang Road, Taichung 402, Taiwan.|Institute of Biomedical Science and Rong Hsing Research Center for Translational Medicine, National Chung Hsing University , No. 250, Guoguang Road, Taichung 402, Taiwan.|Department of Medical Research and Education, Taichung Veterans General Hospital , No. 1650, Sec. 4, Taiwan Boulevard, Taichung 40705, Taiwan.|Division of Chest Medicine, Department of Internal Medicine, Changhua Christian Hospital , No. 480, Zhongzheng Road, Changhua 505, Taiwan.
authors.forename: Chi-Chen
authors.initials: CC
authors.lastname: Lin
authors.affiliation: Institute of Biomedical Science, National Chung-Hsing University , No. 250, Guoguang Road, Taichung 402, Taiwan.|Division of Chest Medicine, Department of Internal Medicine, Changhua Christian Hospital , No. 480, Zhongzheng Road, Changhua 505, Taiwan.
authors.forename: Sheng-Hao
authors.initials: SH
authors.lastname: Lin
authors.affiliation: Division of Chest Medicine, Department of Internal Medicine, Changhua Christian Hospital , No. 480, Zhongzheng Road, Changhua 505, Taiwan.|Department of Respiratory Care, College of Health Sciences, Chang Jung Christian University , No. 1, Changda Road, Tainan 71101, Taiwan.|School of Medicine, Chung Shan Medical University , No. 110, Sec. 1, Jianguo North Road, Taichung 40201, Taiwan.
authors.forename: Ching-Hsiung
authors.initials: CH
authors.lastname: Lin
authors.affiliation: Department of Medical Science and Institute of Bioinformatics and Structural Biology, National Tsing Hua University , No. 101, Kuang-Fu Road Sec. 2, Hsinchu 30013, Taiwan.
authors.forename: Ting-Wen
authors.initials: TW
authors.lastname: Chung
authors.affiliation: Department of Medical Science and Institute of Bioinformatics and Structural Biology, National Tsing Hua University , No. 101, Kuang-Fu Road Sec. 2, Hsinchu 30013, Taiwan.
authors.forename: En-Chi
authors.initials: EC
authors.lastname: Liao
authors.affiliation: Department of Medical Science and Institute of Bioinformatics and Structural Biology, National Tsing Hua University , No. 101, Kuang-Fu Road Sec. 2, Hsinchu 30013, Taiwan.
authors.forename: Ying-Jen
authors.initials: YJ
authors.lastname: Chen
authors.affiliation: Department of Medical Science and Institute of Bioinformatics and Structural Biology, National Tsing Hua University , No. 101, Kuang-Fu Road Sec. 2, Hsinchu 30013, Taiwan.
authors.forename: Yu-Shan
authors.initials: YS
authors.lastname: Wei
authors.affiliation: Department of Medical Science and Institute of Bioinformatics and Structural Biology, National Tsing Hua University , No. 101, Kuang-Fu Road Sec. 2, Hsinchu 30013, Taiwan.
authors.forename: Yi-Ting
authors.initials: YT
authors.lastname: Tsai
authors.affiliation: Department of Medical Science and Institute of Bioinformatics and Structural Biology, National Tsing Hua University , No. 101, Kuang-Fu Road Sec. 2, Hsinchu 30013, Taiwan.
authors.forename: Hong-Lin
authors.initials: HL
authors.lastname: Chan
mesh_terms: D000230:Adenocarcinoma / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D000077192:Adenocarcinoma of Lung; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D037282:Calreticulin / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D038181:FMN Reductase / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008175:Lung Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D000068437:Pemetrexed / Q000494:pharmacology*; D020543:Proteome / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000970:Antineoplastic Agents; C576483:CALR protein, human; D037282:Calreticulin; D009363:Neoplasm Proteins; D020543:Proteome; D034741:RNA, Small Interfering; D000068437:Pemetrexed; D038181:FMN Reductase
keywords: DIGE; lung adenocarcinoma; pemetrexed; proteomics; resistance