Alopecurone B reverses doxorubicin-resistant human osteosarcoma cell line by inhibiting P-glycoprotein and NF-kappa B signaling

Yuan-Zheng Xia; Kai Ni; Chao Guo; Chao Zhang; Ya-Di Geng; Zhen-Dong Wang; Lei Yang; Ling-Yi Kong

doi:10.1016/j.phymed.2014.12.011

Alopecurone B reverses doxorubicin-resistant human osteosarcoma cell line by inhibiting P-glycoprotein and NF-kappa B signaling

Phytomedicine. 2015 Mar 15;22(3):344-51. doi: 10.1016/j.phymed.2014.12.011. Epub 2015 Jan 28.

Authors

Yuan-Zheng Xia¹, Kai Ni¹, Chao Guo¹, Chao Zhang¹, Ya-Di Geng¹, Zhen-Dong Wang¹, Lei Yang², Ling-Yi Kong³

Affiliations

¹ State Key Laboratory of Natural Medicines, Department of Natural Medicinal Chemistry, China Pharmaceutical University, 24 Tong Jia Xiang, NanJing 210009, People's Republic of China.
² State Key Laboratory of Natural Medicines, Department of Natural Medicinal Chemistry, China Pharmaceutical University, 24 Tong Jia Xiang, NanJing 210009, People's Republic of China. Electronic address: dorothy19802003@gmail.com.
³ State Key Laboratory of Natural Medicines, Department of Natural Medicinal Chemistry, China Pharmaceutical University, 24 Tong Jia Xiang, NanJing 210009, People's Republic of China. Electronic address: cpu_lykong@126.com.

PMID: 25837271
DOI: 10.1016/j.phymed.2014.12.011

Abstract

Doxorubicin (DOX) was first used in osteosarcoma in the early 1970s as a first-line antineoplastic drug. However, the occurrence of drug resistance in chemotherapeutic treatment has greatly restricted its use. When resistance to DOX treatment occurs, osteosarcoma may become not only resistant to the drug originally administered but also to a wide variety of structurally and mechanistically unrelated drugs. Thus, there is an urgent need to find ways of reversing DOX chemotherapy resistance in osteosarcoma. Plant-derived agents have great potential in preventing the onset of the carcinogenic process and enhancing the efficacy of conventional antitumor drugs. Alopecurone B (ALOB), a flavonoid, is isolated from Traditional Chinese Medicine Sophora alopecuroides L., and is reported to have potent inhibitory effect on multidrug resistance associated protein 1. In this study, a DOX-resistant osteosarcoma cell line (MG-63/DOX) was established by increasing the concentration gradient of DOX in a stepwise manner. MTT assay, flow cytometry analysis, dual-luciferase reporter gene assay, quantitative real-time polymerase chain reaction and Western blot analysis were applied to investigate the reversing effect of ALOB and its underlying mechanisms. The results indicated that ALOB mediated the resistance of MG-63/DOX cells to DOX by inhibiting P-glycoprotein function, transcription and expression. Besides, ALOB also enhanced the sensitivity of MG-63/DOX cells to other conventional chemotherapeutic drugs. Cell viability assay confirmed the reversing activity of ALOB. Furthermore, ALOB increased DOX-induced apoptosis at nontoxic concentration. In addition, ALOB showed inhibitory effect on NF-κB transcription in a DOX-independent manner. Furthermore, NF-κB signaling was suppressed by ALOB in an IKK-dependent manner. These studies not only demonstrate that ALOB is a potential agent for reversal of drug resistant cancers, but also testify that ALOB reverses multidrug resistance by inhibiting P-glycoprotein via NF-κB signaling.

Keywords: Alopecurone B; Doxorubicin; Flavonoid; Multidrug resistance; Nuclear factor-kappa B; P-glycoprotein.

Publication types

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

MeSH terms

ATP Binding Cassette Transporter, Subfamily B / metabolism
Antineoplastic Agents / pharmacology*
Apoptosis / drug effects
Cell Line, Tumor / drug effects
Doxorubicin / pharmacology
Drug Resistance, Neoplasm / drug effects*
Flavonoids / pharmacology*
Humans
NF-kappa B / metabolism
Osteosarcoma / pathology*
Signal Transduction / drug effects*
Stilbenes / pharmacology*

Substances

ABCB1 protein, human
ATP Binding Cassette Transporter, Subfamily B
Antineoplastic Agents
Flavonoids
NF-kappa B
Stilbenes
alopecurone B
Doxorubicin