Inhibitory effect of 4-chlorobenzoyl berbamine on imatinib-resistant K562 cells in vitro and in vivo.
- Author:
Yun-feng ZHANG
1
;
Gen-bo XU
;
Yi-chao GAN
;
Xiao-hua XU
;
Rong-zhen XU
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Antineoplastic Agents; pharmacology; therapeutic use; Benzamides; Benzylisoquinolines; pharmacology; therapeutic use; Drug Resistance, Neoplasm; Female; Fusion Proteins, bcr-abl; metabolism; Gene Expression Regulation, Neoplastic; Humans; I-kappa B Kinase; metabolism; Imatinib Mesylate; K562 Cells; Liver Neoplasms, Experimental; drug therapy; metabolism; Mice; Mice, Nude; Piperazines; pharmacology; Protein-Tyrosine Kinases; antagonists & inhibitors; Pyrimidines; pharmacology; Transcription Factor RelA; metabolism; Xenograft Model Antitumor Assays
- From: Journal of Southern Medical University 2011;31(12):1997-2001
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo observe the inhibitory effect of 4-chlorobenzoyl berbamine (BBD9) on imatinib-resistant cell line K562 (K562/IR) in vitro and in vivo and explore the mechanisms.
METHODSThe IC50 of BBD9 and berbamine (BBM) was determined by MTT assay. The expressions of p210(Bcr-Abl), IKKa, cytoplasmic and nuclear NF-κBp65 were determined using Western blotting in K562/IR cells following a 48-h exposure to 0.5 µg/ml BBD9 or 8 µg/ml BBM. Flow cytometry was used to analyze the cell viability, apoptosis and necrosis; Western blotting was employed to determine the expressions of PARP, caspase-3, caspase-9 and LC3II in K562/IR cells exposed to different concentrations of BBD9 for 48 h. In nude mouse models bearing K562/IR cell xenograft, the tumor weight, tumor regression, and body weight changes of the mice were measured after treatments with 15 mg/kg and 30 mg/kg BBD9 and 100 mg/kg imatinib.
RESULTSThe IC50 of BBD9 and BBM was 0.73 µg/ml and 5.43 µg/ml, respectively. In K562/IR cell cultures, the expressions of p210(Bcr-Abl), IKKa and nuclear NF-κB p65 were all decreased following BBD9 and BBM treatments, but BBD9 produced more potent effect; cytoplasmic NF-κB p65 showed no obvious changes after the treatments. The cell apoptosis and necrosis increased with the concentrations of BBD9, which also dose-dependently increased the levels of cleaved caspase-3, csapase-9, PARP, and LC3II expression. In the tumor-bearing mouse model, BBD9 showed stronger effects than imatinib in reducing the tumor weight, promoting tumor regression, and increasing the body weight.
CONCLUSIONBBD9 can effectively inhibit the growth of K562/IR cells in vitro and in vivo by activating cell apoptosis, necrosis and autophage pathways, down-regulating expressions of p210(Bcr-Abl) and IKKa and suppressing the cytoplasm-to- nucleus translocation of NF-κBp65.