[Establishment of protein profile of human small cell lung cancer cell line NCI-H446].
OBJECTIVE
Small cell lung cancer (SCLC) is particularly aggressive, and characterized by rapid growth and early metastasis. At present, there is no data concerning SCLC two-dimensional polyacrylamide gel electrophoresis (2-DE) reference map,and its protein profiles in public databases. This study was to establish a well-resolved, reproducible 2-DE map of proteome in SCLC cell line NCI-H446, and analyze its protein profiles.


METHODS
Two-DE was applied to separate the total proteins of NCI-H446 cells, which were then silver-stained in the gel. Well-separated protein spots were selected from the gel by ImageMaster 2D analysis system. Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), peptide map fingerprinting (PMF),and database searching were used to identify the protein spots.


RESULTS
Clear,well-resolved, reproducible 2-DE patterns of proteome in NCI-H446 cells were obtained. The average protein spots of 3 gels were 1506+/-74; and 1412+/-56 spots were matched with an average matching rate of 93.4%. The average deviation of spot position was (0.96+/-0.27) mm in IEF direction, and (1.24+/-0.41) mm in SDS-PAGE direction indicating relatively good reproducibility of the protein spots. Fifty-eight proteins were identified, certain proteins were products of oncogenes, and others were involved in cell cycle regulation, and signal transduction.


CONCLUSIONS
A reference map of NCI-H446 cells was established,certain proteins were identified by MALDI-TOF-MS and PMF. These data will be useful for establishing human SCLC proteome database.
Medical Experimental Research Center,Xiangya Hospital, Central South University, Changsha, Hunan, 410008, P.R.China.
Mao-Yu
MY
Li
Zhi-Qiang
ZQ
Xiao
Cui
C
Li
Xiao-Ying
XY
Wu
Xue-Ping
XP
Feng
Hong
H
Yi
Jian-Ling
JL
Li
Zhu-Chu
ZC
Chen
Ping
P
Chen
Song-Ping
SP
Liang
D000595:Amino Acid Sequence; D018288:Carcinoma, Small Cell / Q000737:chemistry* / Q000473:pathology; D045744:Cell Line, Tumor; D019992:Databases as Topic; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D008175:Lung Neoplasms / Q000737:chemistry* / Q000473:pathology; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000032:analysis* / Q000737:chemistry; D010449:Peptide Mapping; D020543:Proteome; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D009363:Neoplasm Proteins; D020543:Proteome