Differential
Proteomic Analysis of Human Lung Adenocarcinoma Cell Line A-549 and of Normal
Cell Line HBE
ZHAN Xian-Quan, GUAN Yong-Jun, LI Cui,
CHEN Zhu-Chu*
( Cancer Research Institute, Xiangya Medical School, Central South
University, Changsha 410078, China )
XIE Jing-Yun, CHEN Ping, LIANG Song-Ping
( College of Life Science, Hunan Normal University, Changsha 410081,
China )
Abstract To explore
the differential proteomic expressions between human lung adenocarcinoma cell
line A-549 and normal cell line HBE, a series of methods, including immobilized
pH gradient-two dimensional polyacrylamide gel electrophoresis, silver
staining, PDQuest 2-DE software analysis, peptide mass fingerprinting based on
matrix-assisted laser desorption/ionization time of flight mass spectrometry
(MALDI-TOF-MS) and SWISS-PROT database searching, were used to separate and
identify the differential proteomic expressions between A-549 and HBE. The
results showed that the good 2-DE pattern including high resolution and
reproducibility was obtained. After silver staining, the 2-DE image analysis by
PDQuest 2-DE software detected average (890±38) spots in A-549, and (757±27)
spots in HBE. Theaverage positional deviation of the matched spots between
A-549 and HBE 2-DE maps was (2.85±0.48) mm in IEF direction, and (2.69±0.37) mm
in SDS-PAGE direction.The differential proteomic expression analysis found that
there were 535 matched spots between A-549 and HBE 2-DE maps, 355 spots that
were not matched in A-549, 222spots that were not matched in HBE. 18
differential spots (8 spots in A-549 and 10spots in HBE) were cut off from
silver staining gel at random, digested in gel with TPCK-trypsin, measured with
MALDI-TOF-MS and searched in the SWISS-PROT database with PeptIdent software.
18 protein were preliminarily identified. These proteins were related to cell
signal transduction, cell metabolism, proliferation and differentiatiation etc.
There was a significant difference at protein level between human lung adenocarcinoma cell line A-549 and normal
cell line HBE. It suggests that the differential expression analysis of
proteomes may be useful to further study of the related proteins and the
molecular markers of lung adenocarcinoma.
Key words human lung adenocarcinoma cell (A-549); two
dimensional electrophoresis; image analysis; peptide mass fingerprinting
analysis; proteomic comparison
*Corresponding author: Tel,
86-731-4805447; Fax, 86-731-4485482; e-mail, [email protected]