Establishment of a bortezomib-resistant myeloma cell line and differential proteins analysis by MALDI-OF-MS.
- Author:
Rong ZHU
1
;
Hao XI
;
Yong-Hua LI
;
Hua JIANG
;
Jian-Feng ZOU
;
Jian HOU
Author Information
- Publication Type:Journal Article
- MeSH: Antineoplastic Agents; pharmacology; Boronic Acids; pharmacology; Bortezomib; Cell Line, Tumor; Drug Resistance, Neoplasm; genetics; Humans; Multiple Myeloma; pathology; Myeloma Proteins; analysis; Proteome; analysis; Pyrazines; pharmacology; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; methods
- From: Journal of Zhejiang University. Medical sciences 2009;38(5):445-452
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish a bortezomib-resistant myeloma cell line and to investigate its mechanism.
METHODSBortezomib-resistant NCI-H929 cell line (NCI-H929B) was obtained by stepwise increasing extracellular concentrations of bortezomib over a period of 8 months. The biological characteristics of NCI-H929 and NCI-H929B were observed. Proteins from NCI-H929B cell and NCI-H929 cell were extracted, run on two-dimensional gel electrophoresis. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) and mass spectrometry (MS) were used to identify proteins. Western blot was used to further verify differential proteins.
RESULTBortezomib-resistant cell line NCI-H929B was established. NCI-H929B exhibits a 23.5-fold level of resistance to bortezomib as compared to the parental cell line NCI-H929. There were no significant differences in cellular biology of cell growth curve and cell cycle distribution between NCI-H929 and NCI-H929B cell lines.Whole proteins of NCI-H929 and NCI-H929B myeloma cell lines were extracted by two-dimensional gel electrophoresis. Gel-image analysis revealed that there were 17 differential protein spots. A total of 14 differential protein spots were successfully identified by MALDI-TOF-MS. The result of Western blot was consistent with 2-DE.
CONCLUSIONA bortezomib-resistant human myeloma cell line NCI-H929B was successfully established. The differentially expression of proteomes may be useful for study of the bortezomib-resistant mechanisms and the molecular markers of MM.
