期刊论文详细信息
BMC Complementary and Alternative Medicine
Studies on the purification of polypeptide from sika antler plate and activities of antitumor
Xing Y. Meng2  Lei Wu2  Rui Hu3  Yu H. Tian1  Lin Qi2  Wei Hu2 
[1] China Agriculture University, Beijing 100094, China;College of Life Sciences, Jilin Agriculture University, No. 2888 Xincheng Road, Changchun 130118, Jilin province, China;The branch institute of Liaodong college agronomy courtyard, Dandong 118001, China
关键词: Cell proliferation;    Antitumor activities;    Monomer polypeptide;    Antler plate;    Sika;   
Others  :  1226051
DOI  :  10.1186/s12906-015-0845-7
 received in 2014-12-17, accepted in 2015-09-02,  发布年份 2015
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【 摘 要 】

Background

We isolated a novel monomeric peptide from antler plate polypeptide (APP) of sika deer and found that it inhibited rat breast cancer cell proliferation and telomerase activity.

Methods

The molecular mass and purity of this polypeptide was determined by ultra performance liquid chromatography (UPLC) and Bruker micOTOF OllQ TOF mass spectrometry, respectively. The full amino-acid sequence of the monomeric peptide was analyzed by sequential Edman degradation using a protein/peptide sequencer. The APP-1 markedly inhibited rat breast cancer cell proliferation as determined with an 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H- tetrazolium bromide (MTT) assay. Then, we used flow cytometry to detect the effects of the monomeric peptide on cell cycle. Relative quantitative fluorescence PCR was used to analyze the expression level telomerase reverse transcriptase (TERT).

Results

The molecular mass and purity of this polypeptide was 10646 Da and 91.2 %. Amino acid sequence analyses indicated that the N-terminal amino-acid sequence of this monomeric peptide was: MTKLE DYLEG IVNIF HQYSV. The results showed that monomeric peptide halted most cancer cells stagnating in the G0/G1 phase. The percentage of cells in the G 0 /G 1is higher than control group after the monomeric peptide treatment. Relative quantitative fluorescence PCR results showed that TERT gene expression level obviously decreased after treatment with the monomeric peptide compared with control group.

Conclusions

Collectively, the results suggest that this novel and monomeric APP has antitumor activities and imply that it is likely an important component of antitumor activities in antler plate polypeptide.

【 授权许可】

   
2015 Hu et al.

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【 参考文献 】
  • [1]Zhang B. The chemical composition and exploitation of the antler plate. Strain Economic Animals Plants. 2005; 12:7.
  • [2]Wu F, Jin LJ, Li XY. New progress in natural active ingredients with pharmacological functions of deer antler base. Heilongjiang Animal Science and Veterinary Medicine. 2013; 1:23-5.
  • [3]Tang RN. Studies on the Purification and Activities of 17kDa Polypeptide from Sika Antler Plate. Chin Med Science Chang Chun. 2008;4–7.
  • [4]Suh SJ, Kim KS, Lee AR. Prevention of collagen induced arthritis in mice by Cervus Korean TEMMINC K var. Mantchuric us Swinhoe [J]. Enviro Toxico Phar. 2007; 23:147-53.
  • [5]Li C, Yang F, Sheppard A. Adult stem cells and mammalian epimorphic regeneration-insights from studying annual renewal of deer antlers. Curr Stem Cell Res Ther. 2009; 4:237-51.
  • [6]Li C, Yang F, Xing X, Gao X, Deng X, Mackintosh C et al.. Role of heterotypic tissue interactions in deer pedicle and first antler formation-revealed via a membrane insertion approach. J Exp Zool B Mol Dev Evol. 2008; 310:267-77.
  • [7]Kierdorf U, Li C, Price JS. Improbable appendages: Deer antler renewal as a unique case of mammalian regeneration. Semin Cell Dev Biol. 2009; 20:535-42.
  • [8]Kierdorf U, Kierdorf H. Delayed ectopic antler growth and formation of a double-head antler in the metacarpal region of a fallow buck (Dama dama L.) following transplantation of antlerogenic periosteum. Ann Anat. 2000; 182:365-70.
  • [9]Qian J, Ji JX, Huang FJ. Study and Utilization of the Active Substance in Antler Plate. Pharm Biotech. 2009; 16:283-6.
  • [10]Wang ZB, Qiu FP, Li ZM, Zhao LD. Effects of active coronet compos ition on macrophage phagocytos is of mice and hormone level of mas toplas ia in rats. Food Scie Tech. 2007; 11:225-6.
  • [11]Yu H, Wei H. Study and Utilization of the Antler Plate and Its By-product. Lishizhen Medicine and Materia Medica Research. 2010; 21:2310.
  • [12]Huang FJ. Isolation, Characterization and Biological Activity of a Novel Peptide from Sika (Cervus nippon Temminck) Antler. Drugs Biotech. 2010; 17:151-6.
  • [13]Zhang Z, Liu X, Duan L, Li X, Yang Z, Zhou Q. The effects of velvet antler polypeptides on the phenotype and related biological indicators of osteoarthritic rabbit chondrocytes. Biochimica Polonica. 2011; 58:297-302.
  • [14]Guan S-W, Duan L-X, Li Y-Y, Wang B-X, Zhou Q-L. A novel polypeptide from Cervus nippon Temminck proliferation of epidermal cells and NIH3T3 cell line. Acta Biochim Pol. 2006; 53:395-7.
  • [15]Avula B, Wang YH, Pawar RS, Shukla YJ, Smillie TJ, Khan IA. A rapid method for chemical fingerprint analysis of Hoodia species, related genera, and dietary supplements using UPLC-UV-MS. J Pharm Biomed Anal. 2008; 48:722-31.
  • [16]Xinsa FU, Congxiang SHEN, Guanxue LI, Xinyu ZHANG, Zhong WEN. Quantitative detection of plasma level of human telomerase reverse transcriptase mRNA in patients with nasopharyngeal carcinoma. J South Med Univ. 2015; 35:894-7.
  • [17]Nirogi R, Kandikere V, Mudigonda K, Ajjala D. Liquid chromatography tandem mass spectrometry method for the quantification of rimonabant, a CB1 receptor antagonist, in human plasma. Biomed Chroma. 2008; 22:469-77.
  • [18]Hsieh Y, Duncan CJG, Brisson CJG. Fused-core silica column high-performance liquid chromatography/tandem mass spectrometric determination of rimonabant in mouse plasma. Analytical Chem. 2007; 79:5668-73.
  • [19]Xu YL, Yang LJ, Liu GF. Purification of cytotoxin from Naj a naja atra venom and its antitumor activity in vitro. Chin J of Biochemical Phar. 2003; 24:127-30.
  • [20]He DW, Chen H, Ye YY. Study on the anti-tumor effects of earthworm extracts bothin vivoandin vitro. Chin J of Biochemical Phar. 2005; 26:353-5.
  • [21]Jia SQ, Wang WL, Su XL. Inhibitory effect of anti-cancer bioactive peptide on the proliferation of human breast cancer cell line nm231. Chin Med Biotechnol. 2007; 2:270-5.
  • [22]Ling-ping KONG, Hua-qiao WANG. Telomere, Telomerase and Its Potential Relation-ship with Aging, Cancer:A Brief Introduction to the Nobel Prize in Physiology or Medicine 2009. Chin Journal of Nature. 2009; 31:327-31.
  • [23]Didonna S, Mamchaoui K, Cooper R. N. Te-lomerase can extend the proliferative capacity of human myo-blasts, but does not lead to their immortalization. Mol Cancer Res. 2003; 1:643-53.
  • [24]Flores I, Blasco MA. The role of tolemeres and telomerase in stem cell aging. FEBS Lett. 2010; 584:3826-30.
  • [25]Danied M, Peek GW, Tollefsbol TO. Regulation of the human catalytic subunit of telomerase (hTERT). Gene. 2012; 498:135-46.
  • [26]Luo S, Hu YZ, Peng HP. The Effect of Telomerase Activity And Cell Apoptosis on Human Hepatocarcinoma Cell Line (BEL-7402) By Fraction C from Naja Naja Atra Venom. Academic J Guangzhou Med Ical College. 2003; 31:1-6.
  • [27]Li GS, Yang CX, Wei XM, Zhu L. Effect of buthus martensii kirsch on telomere enzymatic activity of liver cancer BelO7404 cell. J Southeast Univ. 2003; 23:166-9.
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