Establishment of two-dimensional gel electrophoresis profiles of the human acute promyelocytic leukemia cell line NB4

He,Pengcheng; Liu,Yanfeng; Zhang,Mei; Wang,Xiaoning; Wang,Huaiyu; Xi,Jieying; Wei,Kaihua; Wang,Hongli; Zhao,Jing

doi:10.3892/mmr.2012.963

Establishment of two-dimensional gel electrophoresis profiles of the human acute promyelocytic leukemia cell line NB4

Authors:
- Pengcheng He
- Yanfeng Liu
- Mei Zhang
- Xiaoning Wang
- Huaiyu Wang
- Jieying Xi
- Kaihua Wei
- Hongli Wang
- Jing Zhao
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Affiliations: Department of Hematology, The First Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an 710061, P.R. China, National Center of Biomedical Analysis, Beijing 100094, P.R. China
Published online on: June 21, 2012 https://doi.org/10.3892/mmr.2012.963
Pages: 570-574

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Abstract

To explore optimum conditions for establishing a two‑dimensional gel electrophoresis (2-DE) map of the human acute promyelocytic leukemia (APL) cell line NB4 and to analyze its protein profiles, we extracted total proteins from NB4 cells using cell disruption, liquid nitrogen freeze-thawing and fracturing by ultrasound, and quantified the extracted protein samples using Bradford's method. 2-DE was applied to separate the proteins, which were silver-stained in the gel. Well‑separated protein spots were selected from the gel using the ImageMaster™ 2D Platinum analysis system. Moreover, the effects of various protein sample sizes (140, 160 and 180 µg) on the 2-DE maps of the NB4 cells were determined and compared. Matrix-assisted laser desorption/ionization time of flight-mass spectrometry (MALDI-TOF-MS), peptide mass fingerprinting (PMF) and database searching were used to identify the proteins. When the quantity of loading proteins was 160 µg, clear, well-resolved, reproducible 2-DE proteomic profiles of the NB4 cells were obtained. The average number of protein spots in 3 gels was 1160±51 with an average matching rate of 81%. A total of 10 proteins were identified by mass spectrometry and database queries, certain proteins were products of oncogenes and others were involved in cell cycle regulation and signal transduction. In summary, 2-DE profiles of the proteome of NB4 cells were established and certain proteins were identified by MALDI-TOF-MS and PMF which lay the foundation of further proteomic research of NB4 cells. These data should be useful for establishing a human APL proteome database.

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