Sensitivity to bortezomib of RPIM8226 cells after co-cultured with down-regulated Cav-1 expression HUVECs.
- Author:
Lu CHEN
1
;
Song-guang JU
;
Zi-yan WANG
;
Jun LI
;
Yu-qing YUAN
;
Jin-xiang FU
Author Information
- Publication Type:Journal Article
- MeSH: Apoptosis; Boronic Acids; pharmacology; Bortezomib; Caveolin 1; metabolism; Cell Line, Tumor; drug effects; Cells, Cultured; Coculture Techniques; Down-Regulation; Human Umbilical Vein Endothelial Cells; cytology; metabolism; Humans; Multiple Myeloma; Pyrazines; pharmacology
- From: Chinese Journal of Hematology 2013;34(11):946-951
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the sensitivity to bortezomib of RPMI8226 cells after co-cultured with down-regulated Caveolin (Cav)-1 expression of HUVECs by transfection with Cav-1 shRNA (HUVECs(Cav-1 low)).
METHODSExposure to bortezomib with or without 50 nmol/L dexamethasone at different concentration, the proliferation of RPMI8226 was analyzed by MTT assay when it was cultured alone or co-cultured with HUVECs(Cav-1 low). Cav-1 expression was detected by using of Western blot and cell cycle, apoptosis and the level of reactive oxygen species (ROS) were analyzed by flow cytometry.
RESULTSCav-1 expression was notably down-regulated in HUVECs(Cav-1 low) (0.2199±0.0288 vs 1.3195±0.2393) (P<0.01). The IC(50) of bortezomib for RPMI8226 cultured alone, co-cultured with HUVECs orHUVECCav- 1 low were 20 nmol/L, 50 nmol/L and 65 nmol/L, respectively. The percentages of G₀/G₁ phase in RPMI8226 cultured alone, co-cultured with HUVECs and HUVECs(Cav-1 low) were 28.49%, 30.41%, and 36.15% respectively. The protection of RPMI 8226 against apoptosis by HUVECs was demonstrated that the apoptosis/death rates were 66.8%, 10.7% and 8.6% in RPMI8226 cultured alone, co-cultured with HUVECs and HUVECs(Cav-1 low) after exposure to 20 nmol/L bortezomib for 24 h. RPMI8226 could induce the oxidative stress of HUVECs before and after co-culture. The ROS level was raised from 15.0% to 35.2% in RPMI8226, from 80.4% to 91.0% in HUVECs, and from 84.6% to 96.8% in HUVECs(Cav-1 low).
CONCLUSIONThe down-regulated Cav-1 expression of HUVECs could promote proliferation and induce apoptosis of RMPI8226 cells, lead to G₀/G₁ phase arrest, and reduce the sensitivity to bortezomib.