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Proteomic analysis of cell lines to identify the irinotecan resistance proteins

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Abstract

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.

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Abbreviations

ACN:

acetonitrile

AKR:

aldo-keto reductase

AR1A1:

aldose reductase

CBB:

Coomassie brilliant blue

CFL1:

Cofilin-1

CRYAB:

alpha-crystallin B chain

2-DE:

two-dimensional gel electrophoresis

DMEM:

Dulbecco’s modified Eagle medium

ELISA:

enzyme-linked immunosorbent assay

5-FU:

5-fluorouracil

HSP27:

heat shock protein 27

IEF:

isoelectric focusing

IPG:

immobilised pH gradient

LoVo cells:

human colon adenocarcinoma

LoVo/irinotecan:

irinotecan-resistant human colon adenocarcinoma

MDR:

multidrug resistance

MS:

mass spectrometry

MTT:

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

PBS:

phosphate buffered saline

Pgp:

P-glycoprotein

PVDF:

polyvinylidene difluoride

Q-TOF:

quadrupole time-of-flight

TBST:

Tris-buffered saline Tween-20

TFA:

trifluoroacetic acid

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Authors and Affiliations

  1. Department of Medical Oncology, Cancer Center, and The State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, No. 37, Guo Xue Xiang, Chengdu, 610041, Sichuan Province, China

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

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  1. Xing-Chen Peng
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  2. Feng-Ming Gong
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  3. Meng Wei
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  4. Xi Chen
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  5. Ye Chen
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  6. Ke Cheng
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  7. Feng Gao
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  8. Feng Xu
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  9. Feng Bi
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  10. Ji-Yan Liu
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Correspondence to Ji-Yan Liu.

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These authors contributed equally to this work.

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Peng, XC., Gong, FM., Wei, M. et al. Proteomic analysis of cell lines to identify the irinotecan resistance proteins. J Biosci 35, 557–564 (2010). https://doi.org/10.1007/s12038-010-0064-9

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  • DOI: https://doi.org/10.1007/s12038-010-0064-9

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