Inducing cell cycle arrest and apoptosis by dimercaptosuccinic acid modified Fe3O4 magnetic nanoparticles combined with nontoxic concentration of bortezomib and gambogic acid in RPMI-8226 cells

Wei Zhang; Lixing Qiao; Xinchao Wang; Ravichandran Senthilkumar; Fei Wang; Baoan Chen

doi:10.2147/IJN.S80795

Inducing cell cycle arrest and apoptosis by dimercaptosuccinic acid modified Fe3O4 magnetic nanoparticles combined with nontoxic concentration of bortezomib and gambogic acid in RPMI-8226 cells

Int J Nanomedicine. 2015 Apr 30:10:3275-89. doi: 10.2147/IJN.S80795. eCollection 2015.

Authors

Wei Zhang¹, Lixing Qiao², Xinchao Wang³, Ravichandran Senthilkumar¹, Fei Wang¹, Baoan Chen⁴

Affiliations

¹ Medical School, Southeast University, Nanjing, People's Republic of China.
² Department of Pediatrics, Zhongda Hospital, Medical School, Southeast University, Nanjing, People's Republic of China.
³ Department of Thyroid and Breast, the Fourth Central Hospital, Tianjin, People's Republic of China.
⁴ Medical School, Southeast University, Nanjing, People's Republic of China ; Department of Hematology and Oncology, Zhongda Hospital, Medical School, Southeast University, Nanjing, People's Republic of China.

Abstract

The purpose of this study was to determine the potential benefits of combination therapy using dimercaptosuccinic acid modified iron oxide (DMSA-Fe3O4) magnetic nanoparticles (MNPs) combined with nontoxic concentration of bortezomib (BTZ) and gambogic acid (GA) on multiple myeloma (MM) RPMI-8226 cells and possible underlying mechanisms. The effects of BTZ-GA-loaded MNP-Fe3O4 (BTZ-GA/MNPs) on cell proliferation were assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,4,-diphenyltetrazolium bromide (MTT) method. Cell cycle and apoptosis were detected using the terminal deoxyribonucleotidyl transferase (TdT)-mediated biotin-16-dUTP nick-end labeling (TUNEL) assay and flow cytometry (FCM). Furthermore, DMSA-Fe3O4 MNPs were characterized in terms of distribution, apoptotic morphology, and cellular uptake by transmission electron microscopy (TEM) and 4,6-diamidino-2-phenylindole (DAPI) staining. Subsequently, the effect of BTZ-GA/MNPs combination on PI3K/Akt activation and apoptotic-related protein were appraised by Western blotting. MTT assay and hematoxylin and eosin (HE) staining were applied to elevate the functions of BTZ-GA/MNPs combination on the tumor xenograft model and tumor necrosis. The results of this study revealed that the majority of MNPs were quasi-spherical and the MNPs taken up by cells were located in the endosome vesicles of cytoplasm. Nontoxic concentration of BTZ-GA/MNPs increased G2/M phase cell cycle arrest and induced apoptosis in RPMI-8226 cells. Furthermore, the combination of BTZ-GA/MNPs activated phosphorylated Akt levels, Caspase-3, and Bax expression, and down-regulated the PI3K and Bcl-2 levels significantly. Meanwhile, the in vivo tumor xenograft model indicated that the treatment of BTZ-GA/MNPs decreased the tumor growth and volume and induced cell apoptosis and necrosis. These findings suggest that chemotherapeutic agents polymerized MNPs-Fe3O4 with GA could serve as a better alternative for targeted therapeutic approaches to treat multiple myeloma.

Keywords: DMSA-Fe3O4; PI3K/Akt; apoptosis; bortezomib; cell cycle; gambogic acid; magnetic nanoparticles.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antineoplastic Agents* / chemistry
Antineoplastic Agents* / toxicity
Apoptosis / drug effects*
Bortezomib* / chemistry
Bortezomib* / toxicity
Cell Cycle Checkpoints / drug effects*
Cell Line, Tumor
Humans
Magnetite Nanoparticles* / chemistry
Magnetite Nanoparticles* / toxicity
Succimer / chemistry*
Xanthones* / chemistry
Xanthones* / pharmacology

Substances

Antineoplastic Agents
Magnetite Nanoparticles
Xanthones
Bortezomib
gambogic acid
Succimer