Antiproliferative and apoptosis-inducing activity of schisandrin B against human glioma cells

Cancer Cell International

Zhi-peng Su¹  Wei-ming Zheng¹  Qi-chuan Zhuge¹  Jin-sen Wu¹  Jiang-long Lu¹  Lin Cai¹  Cheng-de Wang¹  Xiang-he Lu¹  Qun Li¹ 

[1] Department of Neurosurgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China

关键词: Apoptosis;    Cell cycle;    Proliferation;    Schisandrin B;    Glioma cells;   

Others  :  1138570
DOI  :  10.1186/s12935-015-0160-x

 received in 2014-10-27, accepted in 2015-01-07,  发布年份 2015

【 摘 要 】

Background

Malignant glioma is the most devastating and aggressive tumour in the brain and is characterised by high morbidity, high mortality and extremely poor prognosis. The main purpose of the present study was to investigate the effects of schisandrin B (Sch B) on glioma cells both in vitro and in vivo and to explore the possible anticancer mechanism underlying Sch B-induced apoptosis and cell cycle arrest.

Methods

The anti-proliferative ability of Sch B on glioma cells were assessed by MTT and clony formation assays. Flow cytometric analysis was used to detect cell cycle changes. Apoptosis was determined by Hoechst 33342 staining and annexin V/PI double-staining assays. The mitochondrial membrane potential was detected by Rhodamine 123 staining. The in vivo efficacy of Sch B was measured using a U87 xenograft model in nude mice. The expressions of the apoptosis-related and cell cycle-related proteins were analysed by western blot. Student’s t-test was used to compare differences between treated groups and their controls.

Results

We found that Sch B inhibited growth in a dose- and time-dependent manner as assessed by MTT assay. In U87 and U251 cells, the number of clones was strongly suppressed by Sch B. Flow cytometric analysis revealed that Sch B induced cell cycle arrest in glioma cells at the G0/G1 phase. In addition, Sch B induced glioma cell apoptosis and reduced mitochondrial membrane potential (ΔΨm) in a dose-dependent manner. Mechanically, western blot analysis indicated that Sch B induced apoptosis by caspase-3, caspase-9, PARP, and Bcl-2 activation. Moreover, Sch B significantly inhibited tumour growth in vivo following the subcutaneous inoculation of U87 cells in athymic nude mice.

Coclusions

In summary, Sch B can reduce cell proliferation and induce apoptosis in glioma cells and has potential as a novel anti-tumour therapy to treat gliomas.

【 授权许可】

   
2015 Li et al.; licensee BioMed Central.

【 图 表 】

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【 参考文献 】

• [1]Wen PY, Kesari S: Malignant gliomas in adults. N Engl J Med 2008, 359(5):492-507.
• [2]DeAngelis LM: Brain tumors. N Engl J Med 2001, 344(2):114-23.
• [3]Prados MD, Levin V: Biology and treatment of malignant glioma. Semin Oncol 2000, 27(3 Suppl 6):1-10.
• [4]Holland EC: Glioblastoma multiforme: the terminator. Proc Natl Acad Sci U S A 2000, 97(12):6242-4.
• [5]Stupp R, Mason WP, van den Bent MJ, Weller M, Fisher B, Taphoorn MJ, et al.: Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med 2005, 352(10):987-96.
• [6]Stupp R, Hegi ME, Mason WP, van den Bent MJ, Taphoorn MJ, Janzer RC, et al.: Effects of radiotherapy with concomitant and adjuvant temozolomide versus radiotherapy alone on survival in glioblastoma in a randomised phase III study: 5-year analysis of the EORTC-NCIC trial. Lancet Oncol 2009, 10(5):459-66.
• [7]Cheng L, Wu Q, Guryanova OA, Huang Z, Huang Q, Rich JN, et al.: Elevated invasive potential of glioblastoma stem cells. Biochem Biophys Res Commun 2011, 406(4):643-8.
• [8]Kortmann RD, Jeremic B, Weller M, Plasswilm L, Bamberg M. Radiochemotherapy of malignant glioma in adults. Clinical experiences. Strahlentherapie und Onkologie : Organ der Deutschen Rontgengesellschaft [et al.] 2003; 179(4):219–32.
• [9]Liu GT: Pharmacological actions and clinical use of fructus schizandrae. Chin Med J (Engl) 1989, 102(10):740-9.
• [10]Li L, Wang T, Xu ZL, Yu Y, Chen W, Chen F: Effects of schisandrin B on reversing multidrug resistance in human breast cancer cells transfected with mdr1 gene. Zhonghua yi xue za zhi 2005, 85(23):1633-7.
• [11]Liu XN, Zhang CY, Jin XD, Li YZ, Zheng XZ, Li L: Inhibitory effect of schisandrin B on gastric cancer cells in vitro. World J Gastroenterol 2007, 13(48):6506-11.
• [12]Wu YF, Cao MF, Gao YP, Chen F, Wang T, Zumbika EP, et al.: Down-modulation of heat shock protein 70 and up-modulation of Caspase-3 during schisandrin B-induced apoptosis in human hepatoma SMMC-7721 cells. World J Gastroenterol 2004, 10(20):2944-8.
• [13]Li L, Lu Q, Shen Y, Hu X: Schisandrin B enhances doxorubicin-induced apoptosis of cancer cells but not normal cells. Biochem Pharmacol 2006, 71(5):584-95.
• [14]Liu Z, Zhang B, Liu K, Ding Z, Hu X: Schisandrin B attenuates cancer invasion and metastasis via inhibiting epithelial-mesenchymal transition. PLoS One 2012, 7(7):e40480.
• [15]Nishida H, Tatewaki N, Nakajima Y, Magara T, Ko KM, Hamamori Y, et al.: Inhibition of ATR protein kinase activity by schisandrin B in DNA damage response. Nucleic Acids Res 2009, 37(17):5678-89.
• [16]Rodriguez-Vargas JM, Ruiz-Magana MJ, Ruiz-Ruiz C, Majuelos-Melguizo J, Peralta-Leal A, Rodriguez MI, et al.: ROS-induced DNA damage and PARP-1 are required for optimal induction of starvation-induced autophagy. Cell Res 2012, 22(7):1181-98.
• [17]Du L, Mei HF, Yin X, Xing YQ: Delayed growth of glioma by a polysaccharide from Aster tataricus involve upregulation of Bax/Bcl-2 ratio, activation of caspase-3/8/9, and downregulation of the Akt. Tumour Biol 2014, 35(3):1819-25.
• [18]She EX, Hao Z: A novel piperazine derivative potently induces caspase-dependent apoptosis of cancer cells via inhibition of multiple cancer signaling pathways. American journal of translational research 2013, 5(6):622-33.
• [19]Nigg EA: Cyclin-dependent protein kinases: key regulators of the eukaryotic cell cycle. Bioessays 1995, 17(6):471-80.
• [20]Saab R, Bills JL, Miceli AP, Anderson CM, Khoury JD, Fry DW, et al.: Pharmacologic inhibition of cyclin-dependent kinase 4/6 activity arrests proliferation in myoblasts and rhabdomyosarcoma-derived cells. Mol Cancer Ther 2006, 5(5):1299-308.
• [21]Marzec M, Kasprzycka M, Lai R, Gladden AB, Wlodarski P, Tomczak E, et al.: Mantle cell lymphoma cells express predominantly cyclin D1a isoform and are highly sensitive to selective inhibition of CDK4 kinase activity. Blood 2006, 108(5):1744-50.
• [22]Harbour JW, Luo RX, Dei Santi A, Postigo AA, Dean DC: Cdk phosphorylation triggers sequential intramolecular interactions that progressively block Rb functions as cells move through G1. Cell 1999, 98(6):859-69.
• [23]Ezhevsky SA, Ho A, Becker-Hapak M, Davis PK, Dowdy SF: Differential regulation of retinoblastoma tumor suppressor protein by G(1) cyclin-dependent kinase complexes in vivo. Mol Cell Biol 2001, 21(14):4773-84.
• [24]Hinds PW, Mittnacht S, Dulic V, Arnold A, Reed SI, Weinberg RA: Regulation of retinoblastoma protein functions by ectopic expression of human cyclins. Cell 1992, 70(6):993-1006.
• [25]Kang N, Zhang JH, Qiu F, Tashiro S, Onodera S, Ikejima T: Inhibition of EGFR signaling augments oridonin-induced apoptosis in human laryngeal cancer cells via enhancing oxidative stress coincident with activation of both the intrinsic and extrinsic apoptotic pathways. Cancer Lett 2010, 294(2):147-58.
• [26]Strasser A, Cory S, Adams JM: Deciphering the rules of programmed cell death to improve therapy of cancer and other diseases. EMBO J 2011, 30(18):3667-83.
• [27]Spencer SL, Sorger PK: Measuring and modeling apoptosis in single cells. Cell 2011, 144(6):926-39.
• [28]Wang J, Wu A, Xu Y, Liu J, Qian X: M(2)-A induces apoptosis and G(2)-M arrest via inhibiting PI3K/Akt pathway in HL60 cells. Cancer Lett 2009, 283(2):193-202.
• [29]Ewings KE, Wiggins CM, Cook SJ: Bim and the pro-survival Bcl-2 proteins: opposites attract, ERK repels. Cell Cycle 2007, 6(18):2236-40.
• [30]Das A, Banik NL, Patel SJ, Ray SK: Dexamethasone protected human glioblastoma U87MG cells from temozolomide induced apoptosis by maintaining Bax:Bcl-2 ratio and preventing proteolytic activities. Mol Cancer 2004, 3(1):36. BioMed Central Full Text
• [31]Zhang YH, Wu YL, Tashiro S, Onodera S, Ikejima T: Reactive oxygen species contribute to oridonin-induced apoptosis and autophagy in human cervical carcinoma HeLa cells. Acta Pharmacol Sin 2011, 32(10):1266-75.
• [32]Carlson BL, Pokorny JL, Schroeder MA, Sarkaria JN. Establishment, maintenance and in vitro and in vivo applications of primary human glioblastoma multiforme (GBM) xenograft models for translational biology studies and drug discovery. Current protocols in pharmacology/editorial board, SJ Enna 2011; Chapter 14:Unit 14 16.
• [33]Hu YP, Tan ZJ, Wu XS, Liu TY, Jiang L, Bao RF, et al.: Triptolide induces s phase arrest and apoptosis in gallbladder cancer cells. Molecules 2014, 19(2):2612-28.
• [34]Xie SQ, Wang JH, Ma HX, Cheng PF, Zhao J, Wang CJ: Polyamine transporter recognization and antitumor effects of anthracenymethyl homospermidine. Toxicology 2009, 263(2–3):127-33.