[Establishment of a bortezomib-resistant myeloma cell line and differential proteins analysis by MALDI-OF-MS].
To establish a bortezomib-resistant myeloma cell line and to investigate its mechanism.



Bortezomib-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.



Bortezomib-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.



A 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.
Zhu
Rong
R
Department of Hematology, Changzheng Hospital, The Second Military Medical University, Shanghai 200003, China.
Xi
Hao
H
Li
Yong-Hua
YH
Jiang
Hua
H
Zou
Jian-Feng
JF
Hou
Jian
J
D000970:Antineoplastic Agents / Q000494:pharmacology; D001897:Boronic Acids / Q000494:pharmacology*; D000069286:Bortezomib; D045744:Cell Line, Tumor; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D006801:Humans; D009101:Multiple Myeloma / Q000473:pathology*; D009194:Myeloma Proteins / Q000032:analysis*; D020543:Proteome / Q000032:analysis; D011719:Pyrazines / Q000494:pharmacology*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D000970:Antineoplastic Agents; D001897:Boronic Acids; D009194:Myeloma Proteins; D020543:Proteome; D011719:Pyrazines; D000069286:Bortezomib