Abstract
Seleno-short-chain chitosan (SSCC) is a synthesized chitosan derivative. In this study, antitumor activity and underlying mechanism of SSCC on human non-small-cell lung cancer A549 cells were investigated in vitro. The MTT assay showed that SSCC could inhibit cell viability in a dose- and time-dependent manner, and 200 μg/ml SSCC exhibited significantly toxic effects on A549 cells. The cell cycle assay showed that SSCC triggered S phase cell cycle arrest in a dose- and time-dependent manner, which was related to a downregulation of S phase associated cyclin A. The DAPI staining and Annexin V-FITC/PI double staining identified that the SSCC could induce A549 cells apoptosis. Further studies found that SSCC led to the generation of reactive oxygen species (ROS) and the disruption of mitochondrial membrane potential (MMP) by DCFH-DA and Rhodamin 123 staining, respectively. Meanwhile, free radical scavengers N-acetyl-l-cysteine (NAC) pretreatment confirmed that SSCC-induced A549 cells apoptosis was associated with ROS generation. Furthermore, real-time PCR and western blot assay showed that SSCC up-regulated Bax and down-regulated Bcl-2, subsequently incited the release of cytochrome c from mitochondria to cytoplasm, activated the increase of cleaved-caspase 3 and finally induced A549 cells apoptosis in vitro. In general, the present study demonstrated that SSCC induced A549 cells apoptosis via ROS-mediated mitochondrial apoptosis pathway.
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References
Anal AK, Stevens WF, Remunan-Lopez C (2006) Ionotropic cross-linked chitosan microspheres for controlled release of ampicillin. Int J Pharm 312:166–173. doi:10.1016/j.ijpharm.2006.01.043
Chan CK, Supriady H, Goh BH, Kadir HA (2015) Elephantopus scaber induces apoptosis through ROS-dependent mitochondrial signaling pathway in HCT116 human colorectal carcinoma cells. J Ethnopharmacol 168:291–304. doi:10.1016/j.jep.2015.03.072
Chang CC, Hung CM, Yang YR, Lee MJ, Hsu YC (2013) Sulforaphane induced cell cycle arrest in the G2/M phase via the blockade of cyclin B1/CDC2 in human ovarian cancer cells. J Ovarian Res 6:41. doi:10.1186/1757-2215-6-41
Danial NN, Korsmeyer SJ (2004) Cell death: critical control points. Cell 116:205–219. doi:10.1016/S0092-8674(04)00046-7
Decker C, Bianchi C, Decker D, Morel F (2001) Photoinitiated polymerization of vinyl ether-based systems. Prog Org Coat 42:253–266. doi:10.1016/S0300-9440(01)00203-X
Deepagan VG, Kwon S, You DG, Nguyen VQ, Um W, Ko H, Lee H, Jo DG, Kang YM, Park JH (2016) In situ diselenide-crosslinked polymeric micelles for ROS-mediated anticancer drug delivery. Biomaterials 103:56–66. doi:10.1016/j.biomaterials.2016.06.044
Dong BY, Ganther HE, Stewart C (2002) Ip C: identification of molecular targets associated with selenium-induced growth inhibition in human breast cells using cDNA microarrays. Cancer Res 62:708–714
Estevez H, Garcia-Lidon JC, Luque-Garcia JL, Camara C (2014) Effects of chitosan-stabilized selenium nanoparticles on cell proliferation, apoptosis and cell cycle pattern in HepG2 cells: comparison with other selenospecies. Colloid Surf B 122:184–193. doi:10.1016/j.colsurfb.2014.06.062
Fernandes AP, Gandin V (2015) Selenium compounds as therapeutic agents in cancer. BBA-Gen Subj 1850:1642–1660. doi:10.1016/j.bbagen.2014.10.008
He N, Shi X, Zhao Y, Tian L, Wang D, Yang X (2013) Inhibitory effects and molecular mechanisms of selenium-containing tea polysaccharides on human breast cancer MCF-7 cells. J Agric Food Chem 61:579–588. doi:10.1021/jf3036929
Hua P, Sun M, Zhang G, Zhang Y, Tian X, Li X, Zhang X (2015) Cepharanthine induces apoptosis through reactive oxygen species and mitochondrial dysfunction in human non-small-cell lung cancer cells. Biochem Biophys Res Commun 460:136–142. doi:10.1016/j.bbrc.2015.02.131
Jiang G, Liu J, Ren B, Tang Y, Owusu L, Li M, Zhang J, Liu L, Li W (2016) Anti-tumor effects of osthole on ovarian cancer cells in vitro. J Ethnopharmacol 193:368–376. doi:10.1016/j.jep.2016.08.045
Lee JS, Hong EK (2010) Hericium erinaceus enhances doxorubicin-induced apoptosis in human hepatocellular carcinoma cells. Cancer Lett 297:144–154. doi:10.1016/j.canlet.2010.05.006
Lee SO, Nadiminty N, Wu XX, Lou W, Dong Y, Ip C, Onate SA, Gao AC (2005) Selenium disrupts estrogen signaling by altering estrogen receptor expression and ligand binding in human breast cancer cells. Cancer Res 65:3487–3492. doi:10.1158/0008-5472.CAN-04-3267
Lin M, Tang S, Zhang C, Chen H, Huang W, Liu Y, Zhang J (2016) Euphorbia factor L2 induces apoptosis in A549 cells through the mitochondrial pathway. Acta Pharmaceutica Sinica B. doi:10.1016/j.apsb.2016.06.008
Liu A, Song W, Cao D, Liu X, Jia Y (2008) Growth inhibition and apoptosis of human leukemia K562 cells induced by seleno-short-chain chitosan. Method find Exp Clin 30:181–186. doi:10.1358/mf.2008.30.3.1213209
Ma WD, Zou YP, Wang P, Yao XH, Sun Y, Duan MH, Fu YJ, Yu B (2014) Chimaphilin induces apoptosis in human breast cancer MCF-7 cells through a ROS-mediated mitochondrial pathway. Food Chem Toxicol 70:1–8. doi:10.1016/j.fct.2014.04.014
Mieyal JJ, Gallogly MM, Qanungo S, Sabens EA, Shelton MD (2008) Molecular mechanisms and clinical implications of reversible protein S-Glutathionylation. Antioxid Redox Sign 10:1941–1987. doi:10.1089/ars.2008.2089
Renner K, Amberger A, Konwalinka G, Kofler R, Gnaiger E (2003) Changes of mitochondrial respiration, mitochondrial content and cell size after induction of apoptosis in leukemia cells. BBA-Mol Cell Res 1642:115–123. doi:10.1016/s0167-4889(03)00105-8
Rukkijakan T, Ngiwsara L, Lirdprapamongkol K, Svasti J, Phetrak N, Chuawong P (2016) A synthetic 2,3-diarylindole induces cell death via apoptosis and autophagy in A549 lung cancer cells. Bioorg Med Chem Lett 26:2119–2123. doi:10.1016/j.bmcl.2016.03.079
Sarada SKS, Himadri P, Ruma D, Sharma SK, Pauline T, Mrinalini (2008) Selenium protects the hypoxia induced apoptosis in neuroblastoma cells through upregulation of Bcl-2. Brain Res 1209:29–39. doi:10.1016/j.brainres.2008.02.041
Shu G, Yang J, Zhao W, Xu C, Hong Z, Mei Z, Yang X (2014) Kurarinol induces hepatocellular carcinoma cell apoptosis through suppressing cellular signal transducer and activator of transcription 3 signaling. Toxicol Appl Pharmacol 281:157–165. doi:10.1016/j.taap.2014.06.021
Simon HU, Hajyehia A, Levischaffer SF (2000) Role of reactive oxygen species (ROS) in apoptosis induction. Apoptosis 5:415–418
Stewart MS, Spallholz JE, Neldner KH, Pence BC (1999) Selenium compounds have disparate abilities to impose oxidative stress and induce apoptosis. Free Radic Biol Med 26:42–48. doi:10.1016/S0891-5849(98)00147-6
Su JY, Luo X, Zhang XJ (2015) Immunosupressive activity of pogostone on T cells: blocking proliferation via S phase arrest. Int Immunopharmacol 26:328–337. doi:10.1016/j.intimp.2015.04.019
Sui Y, Li S, Shi P, Wu Y, Li Y, Chen W, Huang L, Yao H, Lin X (2016) Ethyl acetate extract from Selaginella doederleinii Hieron inhibits the growth of human lung cancer cells A549 via caspase-dependent apoptosis pathway. J Ethnopharmacol 190:261–271. doi:10.1016/j.jep.2016.06.029
Tedeschi H (1980) The mitochondrial membrane potential. Biol Rev 55:171–206. doi:10.1111/j.1469-185X.1980.tb00692.x
Wang J, Li Q, Bao A, Liu X, Zeng J, Yang X, Yao J, Zhang J, Lei Z (2016) Synthesis of selenium-containing Artemisia sphaerocephala polysaccharides: solution conformation and anti-tumor activities in vitro. Carbohydr Polym 152:70–78. doi:10.1016/j.carbpol.2016.06.090
Wu CC, Chan ML, Chen WY, Tsai CY, Chang FR, Wu YC (2005) Pristimerin induces caspase-dependent apoptosis in MDA-MB-231 cells via direct effects on mitochondria. Mol Cancer Ther 4:1277–1285. doi:10.1158/1535-7163.MCT-05-0027
Yang M, Wang K, Gao L, Han YT, Lu JF, Zou TT (1992) Exploration for a natural selenium supplement-characterization and bioactivities of Se-containing polysaccharide from garlic. J Chin Pharm Sci 1:28–32
Zhang CZ, Zhang H, Yun J, Chen GG, Lai PBS (2012) Dihydroartemisinin exhibits antitumor activity toward hepatocellular carcinoma in vitro and in vivo. Biochem Pharmacol 83:1278–1289. doi:10.1016/j.bcp.2012.02.002
Zhang CG, Huang JC, Liu T, Li XY (2015) Anticancer effects of bishydroxycoumarin are mediated through apoptosis induction, cell migration inhibition and cell cycle arrest in human glioma cells. Original Artic 20:1592–1600
Zhang J, Liang Y, Lin Y, Liu Y, You Y, Yin W (2016) IRE1α-TRAF2-ASK1 pathway is involved in CSTMP-induced apoptosis and ER stress in human non-small cell lung cancer A549 cells. Biomed Pharmacother 82:281–289. doi:10.1016/j.biopha.2016.04.050
Zhou Y, Lu N, Zhang H, Wei L, Tao L, Dai Q, Zhao L, Lin B, Ding Q, Guo Q (2013) HQS-3, a newly synthesized flavonoid, possesses potent anti-tumor effect in vivo and in vitro. Eur J Pharm Sci 49:649–658. doi:10.1016/j.ejps.2013.04.016
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Zhao, Y., Zhang, S., Wang, P. et al. Seleno-short-chain chitosan induces apoptosis in human non-small-cell lung cancer A549 cells through ROS-mediated mitochondrial pathway. Cytotechnology 69, 851–863 (2017). https://doi.org/10.1007/s10616-017-0098-z
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DOI: https://doi.org/10.1007/s10616-017-0098-z