A selenosemicarbazone complex with copper efficiently down-regulates the 90-kDa heat shock protein HSP90AA1 and its client proteins in cancer cells

BMC Cancer

Hongtao Shen²  Haichuan Zhu⁵  Mowei Song²  Yonglu Tian⁴  Yafei Huang¹  Hui Zheng³  Ruiyuan Cao¹  Jian Lin⁵  Zhenggang Bi²  Wu Zhong¹ 

[1] Laboratory of Computer-Aided Drug Design & Discovery, Beijing Institute of Pharmacology and Toxicology, Beijing, China

[2] The First Affiliated Hospital of Harbin Medical University, Harbin, China

[3] Center for Human Disease Genomics, Peking University, Beijing, China

[4] Laboratory Animal Centre, Peking University, Beijing, China

[5] Synthetic and Functional Biomolecules Center, College of Chemistry and Molecular Engineering, Peking University, Beijing, China

关键词: AKT1;    PIM1;    HSP90AA1 protein;    RNA-seq;    Oxidative stress;    Cell death;    Selenosemicarbazone;   

Others  :  1121239
DOI  :  10.1186/1471-2407-14-629

 received in 2014-06-27, accepted in 2014-08-20,  发布年份 2014

【 摘 要 】

Background

The 90-kDa heat shock protein HSP90AA1 is critical for the stability of several proteins that are important for tumor progression and thus, is a promising target for cancer therapy. Selenosemicarbazone metal complexes have been shown to possess anticancer activity through an unknown molecular mechanism.

Methods

The MTT assay, fluorescence-activated cell sorting, and fluorescent microscopy were used to analyze the mechanism of the anti-cancer activity of the selenosemicarbazone metal complexes. Additionally, RNA-seq was applied to identify transcriptional gene changes, and in turn, the signaling pathways involved in the process of 2-24a/Cu-induced cell death. Last, the expression of HSP90AA1, HSPA1A, PIM1, and AKT proteins in 2-24a/Cu-treated cells were investigated by western blot analysis.

Results

A novel selenosemicarbazone copper complex (2-24a/Cu) efficiently induced G2/M arrest and was cytotoxic in cancer cells. 2-24a/Cu significantly induced oxidative stress in cancer cells. Interestingly, although RNA-seq revealed that the transcription of HSP90AA1 was increased in 2-24a/Cu-treated cells, western blotting showed that the expression of HSP90AA1 protein was significantly decreased in these cells. Furthermore, down-regulation of HSP90AA1 led to the degradation of its client proteins (PIM1 and AKT1), which are also cancer therapy targets.

Conclusion

Our results showed that 2-24a/Cu efficiently generates oxidative stress and down-regulates HSP90AA1 and its client proteins (PIM1, AKT1) in U2os and HeLa cells. These results demonstrate the potential application of this novel copper complex in cancer therapy.

【 授权许可】

   
2014 Shen et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

【 参考文献 】

• [1]Taipale M, Jarosz DF, Lindquist S: HSP90 at the hub of protein homeostasis: emerging mechanistic insights. Nat Rev Mol Cell Biol 2010, 11(7):515-528.
• [2]Whitesell L, Lindquist SL: HSP90 and the chaperoning of cancer. Nat Rev Cancer 2005, 5(10):761-772.
• [3]Neckers L, Workman P: Hsp90 molecular chaperone inhibitors: are we there yet? Clin Cancer Res: Off J Am Assoc Cancer Res 2012, 18(1):64-76.
• [4]Newman B, Liu Y, Lee HF, Sun D, Wang Y: HSP90 inhibitor 17-AAG selectively eradicates lymphoma stem cells. Cancer Res 2012, 72(17):4551-4561.
• [5]McCleese JK, Bear MD, Fossey SL, Mihalek RM, Foley KP, Ying W, Barsoum J, London CA: The novel HSP90 inhibitor STA-1474 exhibits biologic activity against osteosarcoma cell lines. Inter J Cancer J Inter du Cancer 2009, 125(12):2792-2801.
• [6]Okawa Y, Hideshima T, Steed P, Vallet S, Hall S, Huang K, Rice J, Barabasz A, Foley B, Ikeda H, Raje N, Kiziltepe T, Yasui H, Enatsu S, Anderson KC: SNX-2112, a selective Hsp90 inhibitor, potently inhibits tumor cell growth, angiogenesis, and osteoclastogenesis in multiple myeloma and other hematologic tumors by abrogating signaling via Akt and ERK. Blood 2009, 113(4):846-855.
• [7]Karkoulis PK, Stravopodis DJ, Margaritis LH, Voutsinas GE: 17-Allylamino-17-demethoxygeldanamycin induces downregulation of critical Hsp90 protein clients and results in cell cycle arrest and apoptosis of human urinary bladder cancer cells. BMC Cancer 2010, 10:481. BioMed Central Full Text
• [8]Liu KS, Liu H, Qi JH, Liu QY, Liu Z, Xia M, Xing GW, Wang SX, Wang YF: SNX-2112, an Hsp90 inhibitor, induces apoptosis and autophagy via degradation of Hsp90 client proteins in human melanoma A-375 cells. Cancer Lett 2012, 318(2):180-188.
• [9]Tisato F, Marzano C, Porchia M, Pellei M, Santini C: Copper in diseases and treatments, and copper-based anticancer strategies. Med Res Rev 2010, 30(4):708-749.
• [10]Duncan C, White AR: Copper complexes as therapeutic agents. Metallomics: Integ Bio Sci 2012, 4(2):127-138.
• [11]Hancock CN, Stockwin LH, Han B, Divelbiss RD, Jun JH, Malhotra SV, Hollingshead MG, Newton DL: A copper chelate of thiosemicarbazone NSC 689534 induces oxidative/ER stress and inhibits tumor growth in vitro and in vivo. Free Radic Biol Med 2011, 50(1):110-121.
• [12]Beck R, Verrax J, Gonze T, Zappone M, Pedrosa RC, Taper H, Feron O, Calderon PB: Hsp90 cleavage by an oxidative stress leads to its client proteins degradation and cancer cell death. Biochem Pharmacol 2009, 77(3):375-383.
• [13]Rayman MP: Selenium in cancer prevention: a review of the evidence and mechanism of action. Proc Nutrition Soc 2005, 64(4):527-542.
• [14]Sanmartin C, Plano D, Sharma AK, Palop JA: Selenium compounds, apoptosis and other types of cell death: an overview for cancer therapy. Int J Mol Sci 2012, 13(8):9649-9672.
• [15]Srdic-Rajic T, Zec M, Todorovic T, Andelkovic K, Radulovic S: Non-substituted N-heteroaromatic selenosemicarbazone metal complexes induce apoptosis in cancer cells via activation of mitochondrial pathway. Eur J Med Chem 2011, 46(9):3734-3747.
• [16]Zec M, Srdic-Rajic T, Konic-Ristic A, Todorovic T, Andjelkovic K, Filipovic-Ljeskovic I, Radulovic S: Anti-metastatic and anti-angiogenic properties of potential new anti-cancer drugs based on metal complexes of selenosemicarbazones. Anti Cancer Agents Med Chem 2012, 12(9):1071-1080.
• [17]Tian S, Lin J, Zhou J, Wang X, Li Y, Ren X, Yu W, Zhong W, Xiao J, Sheng F, Chen Y, Jin C, Li S, Zheng Z, Xia B: Beclin 1-independent autophagy induced by a Bcl-XL/Bcl-2 targeting compound, Z18. Autophagy 2010, 6(8):1032-1041.
• [18]Kalinowski DS, Yu Y, Sharpe PC, Islam M, Liao YT, Lovejoy DB, Kumar N, Bernhardt PV, Richardson DR: Design, synthesis, and characterization of novel iron chelators: structure-activity relationships of the 2-benzoylpyridine thiosemicarbazone series and their 3-nitrobenzoyl analogues as potent antitumor agents. J Med Chem 2007, 50(15):3716-3729.
• [19]Lovejoy DB, Sharp DM, Seebacher N, Obeidy P, Prichard T, Stefani C, Basha MT, Sharpe PC, Jansson PJ, Kalinowski DS, Bernhardt PV, Richardson DR: Novel second-generation di-2-pyridylketone thiosemicarbazones show synergism with standard chemotherapeutics and demonstrate potent activity against lung cancer xenografts after oral and intravenous administration in vivo. J Med Chem 2012, 55(16):7230-7244.
• [20]Lovejoy DB, Jansson PJ, Brunk UT, Wong J, Ponka P, Richardson DR: Antitumor activity of metal-chelating compound Dp44mT is mediated by formation of a redox-active copper complex that accumulates in lysosomes. Cancer Res 2011, 71(17):5871-5880.
• [21]Narlik-Grassow M, Blanco-Aparicio C, Cecilia Y, Peregrina S, Garcia-Serelde B, Munoz-Galvan S, Canamero M, Carnero A: The essential role of PIM kinases in sarcoma growth and bone invasion. Carcinogenesis 2012, 33(8):1479-1486.
• [22]Amaravadi R, Thompson CB: The survival kinases Akt and Pim as potential pharmacological targets. J Clin Invest 2005, 115(10):2618-2624.
• [23]Zhang XY, Qiao H, Ni JM, Shi YB, Qiang Y: Preparation of isoliquiritigenin-loaded nanostructured lipid carrier and the in vivo evaluation in tumor-bearing mice. Eur J Pharm Sci 2013, 49(3):411-22.
• [24]Jungwirth U, Kowol CR, Keppler BK, Hartinger CG, Berger W, Heffeter P: Anticancer activity of metal complexes: involvement of redox processes. Antioxidants Redox Signaling 2011, 15(4):1085-1127.
• [25]Bjelogrlic S, Todorovic T, Bacchi A, Zec M, Sladic D, Srdic-Rajic T, Radanovic D, Radulovic S, Pelizzi G, Andelkovic K: Synthesis, structure and characterization of novel Cd(II) and Zn(II) complexes with the condensation product of 2-formylpyridine and selenosemicarbazide Antiproliferative activity of the synthesized complexes and related selenosemicarbazone complexes. J Inorg Biochem 2010, 104(6):673-682.
• [26]Agrawal KC, Booth BA, Michaud RL, Moore EC, Sartorelli AC: Comparative studies of the antineoplastic activity of 5-hydroxy-2-formylpyridine thiosemicarbazone and its seleno-semicarbazone, guanylhydrazone and semicarbazone analogs. Biochem Pharmacol 1974, 23(17):2421-2429.
• [27]Kowol CR, Eichinger R, Jakupec MA, Galanski M, Arion VB, Keppler BK: Effect of metal ion complexation and chalcogen donor identity on the antiproliferative activity of 2-acetylpyridine N, N-dimethyl(chalcogen)semicarbazones. J Inorg Biochem 2007, 101(11–12):1946-1957.
• [28]Gligorijevic N, Todorovic T, Radulovic S, Sladic D, Filipovic N, Godevac D, Jeremic D, Andelkovic K: Synthesis and characterization of new Pt(II) and Pd(II) complexes with 2-quinolinecarboxaldehyde selenosemicarbazone: cytotoxic activity evaluation of Cd(II), Zn(II), Ni(II), Pt(II) and Pd(II) complexes with heteroaromatic selenosemicarbazones. Eur J Med Chem 2009, 44(4):1623-1629.
• [29]Garcia-Carbonero R, Carnero A, Paz-Ares L: Inhibition of HSP90 molecular chaperones: moving into the clinic. Lancet Oncol 2013, 14(9):e358-369.
• [30]Shay KP, Wang Z, Xing PX, McKenzie IF, Magnuson NS: Pim-1 kinase stability is regulated by heat shock proteins and the ubiquitin-proteasome pathway. Mole Cancer Res: MCR 2005, 3(3):170-181.
• [31]Bachmann M, Moroy T: The serine/threonine kinase Pim-1. Int J Biochem Cell Biol 2005, 37(4):726-730.
• [32]Karar J, Maity A: PI3K/AKT/mTOR pathway in angiogenesis. Front Mol Neurosci 2011, 4:51.