Toosendanin mediates cisplatin sensitization through targeting Annexin A4/ATP7A in non-small cell lung cancer cells
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Cisplatin (CDDP) is used in the treatment of non-small cell lung cancer (NSCLC), but due to the development of resistance, the benefit has been limited. Toosendanin (TSN) has shown therapeutic effects on NSCLC; however, the role of TSN on CDDP sensitization in NSCLC remains unknown. The antitumor effects of TSN and CDDP sensitization mediated by TSN were explored. TSN was added in various amounts to measure dose- and time-dependent cytotoxicity. Intracellular CDDP was detected by high-performance liquid chromatography. The protein levels of ATP7A, ATP7B, hCTR1, MRP-2, P-gp and Annexin A4 (Anxa4) were analyzed. The tests were conducted using normal NSCLC (A549 cell line) and CDDP-resistant cells (A549/DDP cell line). Anxa4 promotes CDDP resistance by regulating ATP7A, so Anxa4 was overexpressed and silenced and also transfected with pcMV6 or siRNA/ATP7A, respectively. Mechanistic investigations revealed that TSN decreased relative viability in NSCLC cells. Remarkably, TSN significantly enhanced CDDPsensitization in invalid doses. TSN downregulated Anxa4 expression, enhanced intracellular CDDP, and had no effect on MRP-2, P-gp, ATP7A, ATP7B or hCTR1. Subsequently, overexpression of Anxa4 led to a significant decrease in intracellular CDDP concentration. The adjustment of CDDP concentration regulated by TSN disappeared in Anxa4 or ATP7A-silenced cells. TSN also enhanced CDDP sensitization in single ATP7A-overexpressing cells, but had no effect on cells with simultaneous ATP7A overexpression and Anxa4 silencing. The present study suggests that TSN can mediate CDDP sensitization in NSCLC through downregulation of Anxa4.
KeywordsToosendanin Non-small cell lung cancer cells Cisplatin sensitization Annexin A4
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- 3.Zornosa C, Vandergrift JL, Kalemkerian GP, Ettinger DS, Rabin MS, Reid M, Otterson GA, Koczywas M, D’Amico TA, Niland JC, Mamet R, Pisters KM (2012) First-line systemic therapy practice patterns and concordance with NCCN guidelines for patients diagnosed with metastatic NSCLC treated at NCCN institutions. J Natl Compr Canc Netw 10(7):847–856CrossRefPubMedGoogle Scholar
- 10.Tang MZ, Wang ZF, Shi YL (2004) A study of ultrasound-assisted extraction of toosendanin with water from bark of melia toosendan sieb. T zucc. J Southwest Univ Natl (Nat Sci Ed, Chin) 30(8):714–717Google Scholar
- 11.Tang MZ, Wang ZF, Shi YL (2004) Involvement of cytochrome c release and caspase activation in toosendanin-induced PC12 cell apoptosis. Toxicology 201(1–3):131–138Google Scholar
- 15.Harrington CF, Le Pla RC, Jones GD, Thomas AL, Farmer PB (2010) Determination of cisplatin 1,2-intrastrand guanine-guanine DNA adducts in human leukocytes by high-performance liquid chromatography coupled to inductively coupled plasma mass spectrometry. Chem Res Toxicol 23(8):1313–1321CrossRefPubMedGoogle Scholar
- 20.Li X, Huang JM, Wang JN, Xiong XK, Yang XF, Zou F (2015) Combination of chrysin and cisplatin promotes the apoptosis of HepG2 cells by up-regulating p53. Chem Biol Interact 232:212–220Google Scholar
- 27.Kim A, Enomoto T, Serada S, Ueda Y, Takahashi T, Ripley B, Miyatake T, Fujita M, Lee CM, Morimoto K, Fujimoto M, Kimura T, Naka T (2009) Enhanced expression of Annexin A4 in clear cell carcinoma of the ovary and its association with chemoresistance to carboplatin. Int J Cancer 125(10):2316–2322CrossRefPubMedGoogle Scholar
- 28.Matsuzaki S, Enomoto T, Serada S, Yoshino K, Nagamori S, Morimoto A, Yokoyama T, Kim A, Kimura T, Ueda Y, Fujita M, Fujimoto M, Kanai Y, Kimura T, Naka T (2014) Annexin A4-conferred platinum resistance is mediated by the copper transporter ATP7A. Int J Cancer 134(8):1796–1809CrossRefPubMedGoogle Scholar