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
Article

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.

Keywords

Colon cancer 2-DE drug resistance irinotecan proteomics 

Abbreviations used

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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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

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