Journal of Biosciences

, Volume 35, Issue 4, pp 557–564 | Cite as

Proteomic analysis of cell lines to identify the irinotecan resistance proteins

  • Xing-Chen Peng
  • Feng-Ming Gong
  • Meng Wei
  • Xi Chen
  • Ye Chen
  • Ke Cheng
  • Feng Gao
  • Feng Xu
  • Feng Bi
  • Ji-Yan Liu


Chemotherapeutic drug resistance is a frequent cause of treatment failure in colon cancer patients. Several mechanisms have been implicated in drug resistance. However, they are not sufficient to exhaustively account for this resistance emergence. In this study, two-dimensional gel electrophoresis (2-DE) and the PDQuest software analysis were applied to compare the differential expression of irinotecan-resistance-associated protein in human colon adenocarcinoma LoVo cells and irinotecan-resistant LoVo cells (LoVo/irinotecan). The differential protein dots were excised and analysed by ESI-Q-TOF mass spectrometry (MS). Fifteen proteins were identified, including eight proteins with decreased expression and seven proteins with increased expression. The identified known proteins included those that function in diverse biological processes such as cellular transcription, cell apoptosis, electron transport/redox regulation, cell proliferation/differentiation and retinol metabolism pathways. Identification of such proteins could allow improved understanding of the mechanisms leading to the acquisition of chemoresistance.


Colon cancer 2-DE drug resistance irinotecan proteomics 

Abbreviations used




aldo-keto reductase


aldose reductase


Coomassie brilliant blue




alpha-crystallin B chain


two-dimensional gel electrophoresis


Dulbecco’s modified Eagle medium


enzyme-linked immunosorbent assay




heat shock protein 27


isoelectric focusing


immobilised pH gradient

LoVo cells

human colon adenocarcinoma


irinotecan-resistant human colon adenocarcinoma


multidrug resistance


mass spectrometry


3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide


phosphate buffered saline




polyvinylidene difluoride


quadrupole time-of-flight


Tris-buffered saline Tween-20


trifluoroacetic acid


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Arrigo A P, Simon S, Gibert B, Kretz-Remy C, Nivon M, Czekalla A, Guillet D, Moulin M, Diaz-Latoud C and Vicart P 2007 Hsp27 (HspB1) and alphaB-crystallin (HspB5) as therapeutic targets; FEBS Lett. 581 3665–3674CrossRefPubMedGoogle Scholar
  2. Chen R, Pan S, Brentnall T A and Aebersold R 2005 Proteomic profiling of pancreatic cancer for biomarker discovery; Mol. Cell Proteomics 4 523–533CrossRefPubMedGoogle Scholar
  3. Flagiello D, Apiou F, Gibaud A, Poupon M F, Dutrillaux B and Malfoy B 1997 Assignment of the genes for cellular retinoic acid binding protein 1 (CRABP1) and 2 (CRABP2) to human chromosome band 15q24 and 1q21.3, respectively, by in situ hybridization; Cytogenet. Cell Genet. 76 17–18CrossRefPubMedGoogle Scholar
  4. Forrest G L and Gonzalez B 2000 Carbonyl reductase; Chem. Biol. Interact. 129 2140CrossRefGoogle Scholar
  5. Goldberg R M 2006 Therapy for metastatic colorectal cancer; Oncologist 11 981–987CrossRefPubMedGoogle Scholar
  6. Hütter G and Sinha P 2001 Proteomics for studying cancer cells and the development of chemoresistance; Proteomics 1 1233–1248CrossRefPubMedGoogle Scholar
  7. Jemal A, Siegel R, Ward E, Hao Y, Xu J and Thun M J 2009 Cancer statistics, 2009; CA-Cancer J. Clin. 59 225–249CrossRefPubMedGoogle Scholar
  8. Jin Y and Penning T M 2007 Aldo-keto reductases and bioactivation/detoxication; Annu. Rev. Pharmacol. Toxicol. 47 263–292CrossRefPubMedGoogle Scholar
  9. Judson L K, William P H and Adrience C S 2004 Molecular analysis of alpha B-crystallin in human malignant glioma cell populations; AACR Meeting #2263Google Scholar
  10. Kelly H and Goldberg R M 2005 Systemic therapy for metastatic colorectal cancer: current options, current evidence; J. Clin. Oncol. 23 4553–4560CrossRefPubMedGoogle Scholar
  11. Kuffel M J, Reid J M and Ames M M 1992 Anthracyclines and their C-13 alcohol metabolites: growth inhibition and DNA damage following incubation with human tumor cells in culture; Cancer Chemother. Pharmacol. 30 51–57CrossRefPubMedGoogle Scholar
  12. Lage H 2003 ABC-transporters: implications on drug resistance from microorganisms to human cancers; Int. J. Antimicrob. Agents 22 188–199CrossRefPubMedGoogle Scholar
  13. Li X, Cusack B J, Boucek Jr R J, Mushlin P S, Bledsoe T B, Brenner D E and Olson R D 1991 Effects of daunorubicin and its primary metabolite, daunorubicinol, on cardiac contractility and calcium loading of sarcoplasmic reticulum; FASEB J. 5 A1395Google Scholar
  14. Licata S, Saponiero A, Mordente A and Minotti G 2000 Doxorubicin metabolism and toxicity in human myocardium: role of cytoplasmic deglycosidation and carbonyl reduction; Chem. Res. Toxicol. 13 414–420CrossRefPubMedGoogle Scholar
  15. Ling V 1997 Multidrug resistance: molecular mechanisms and clinical relevance; Cancer Chemother. Pharmacol. Suppl. 40 S 38CrossRefGoogle Scholar
  16. Mariann P, Michael S, Christoph H, Lutz K and Edmund M 2007 Increased resistance of tumor cells to daunorubicin after transfection of CDNAs coding for anthracycline inactivating enzymes; Cancer Lett. 255 49–56CrossRefGoogle Scholar
  17. Matsunaga T, Shintani S and Hara A 2006 Multiplicity of mammalian reductases for xenobiotic carbonyl compounds; Drug Metab. Pharmacokinet. 21 118CrossRefGoogle Scholar
  18. Mordente A, Minotti G, Martorana G E, Silvestrini A, Giardina B and Meucci E 2003 Anthracycline secondary alcohol metabolite formation in human or rabbit heart: biochemical aspects and pharmacologic implications; Biochem. Pharmacol. 66 989–998CrossRefPubMedGoogle Scholar
  19. Ohara H, Miyabe Y, Deyashiki Y, Matsuura K and Hara A 1995 Reduction of drug ketones by dihydrodiol dehydrogenases, carbonyl reductase and aldehyde reductase of human liver; Biochem. Pharmacol. 50 221–227CrossRefPubMedGoogle Scholar
  20. Olson R D, Mushlin P S, Brenner D E, Fleischer S, Cusack B J, Chang B K and Boucek Jr R J 1988 Doxorubicin cardiotoxicity may be caused by its metabolite, doxorubicinol; Proc. Natl. Acad. Sci. USA 85 3585–3589CrossRefPubMedGoogle Scholar
  21. Perez R P 1998 Cellular and molecular determinants of cisplatin resistance; Eur. J. Cancer 34 1535–1542CrossRefPubMedGoogle Scholar
  22. Pommier Y, Pourquier P, Fan Y and Strumberg D 1998 Mechanism of action of eukaryotic DNA topoisomerase I and drugs targeted to the enzyme; Biochim. Biophys. Acta 1400 83–105PubMedGoogle Scholar
  23. Schott B and Robert J 1989 Comparative activity of anthracycline 13-hydroxymetabolites against rat glioblastoma cells in culture; Biochem. Pharmacol. 38 4069–4074CrossRefPubMedGoogle Scholar
  24. Sinha P, Hütter G, Kottgen E, Dietel M, Schadendorf D and Lage H 1999 Increased expression of epidermal fatty acid binding protein, cofilin, and 14-3-3-σ(stratifin) detected by two-dimensional gel electrophoresis, mass spectrometry and microsequencing of drug-resistant human adenocarcinoma of the pancreas; Electrophoresis 20 2952–2960CrossRefPubMedGoogle Scholar
  25. Slatter J G, Su P, Sams J P, Schaaf L J and Wienkers L C 1997 Bioactivation of the anticancer agent CPT-11 to SN-38 by human hepatic microsomal carboxylesterases and the in vitro assessment of potential drug interactions; Drug Metab. Dispos. 25 1157–1164PubMedGoogle Scholar
  26. Tanaka K, Imoto I, Inoue J, Kozaki K, Tsuda H, Shimada Y, Aiko S, Yoshizumi Y, Iwai T, Kawano T and Inazawa J 2007 Frequent methylation-associated silencing of a candidate tumorsuppressor, CRABP1, in esophageal squamous-cell carcinoma; Oncogene 26 6456–6468CrossRefPubMedGoogle Scholar
  27. Tong A, Zhang H, Li Z, Gou L, Wang Z, Wei H, Tang M, Liang S, Chen L, Huang C and Wei Y 2007 Proteomic analysis of liver cancer cells treated with suberonylanilide hydroxamic acid; Cancer Chemother. Pharmacol. 61 791–802CrossRefPubMedGoogle Scholar
  28. Vimalachandran D and Costello E 2004 Proteomic technologies and their application to pancreatic cancer; Expert Rev. Proteomics 1 493–501CrossRefPubMedGoogle Scholar
  29. Watanabe G, Kato S, Nakata H, Ishida T, Ohuchi N and Ishioka C 2009 alphaB-crystallin: a novel p53-target gene required for p53-dependent apoptosis; Cancer Sci. 100 2368–2375CrossRefPubMedGoogle Scholar
  30. Yu D H 1998 The role of oncogenes in drug resistance; Cytotechnology 27 283–292CrossRefPubMedGoogle Scholar
  31. Zeng H, Liu G, Rea P A and Kruh G D 2000 Transport of amphipathic anions by human multidrug resistance protein 3; Cancer Res. 60 4779–4784PubMedGoogle Scholar

Copyright information

© Indian Academy of Sciences 2010

Authors and Affiliations

  • Xing-Chen Peng
    • 1
  • Feng-Ming Gong
    • 1
  • Meng Wei
    • 1
  • Xi Chen
    • 1
  • Ye Chen
    • 1
  • Ke Cheng
    • 1
  • Feng Gao
    • 1
  • Feng Xu
    • 1
  • Feng Bi
    • 1
  • Ji-Yan Liu
    • 1
  1. 1.Department of Medical Oncology, Cancer Center, and The State Key Laboratory of Biotherapy, West China Hospital, West China Medical SchoolSichuan UniversityChengduSichuan Province, China

Personalised recommendations