title: Studies on the purification of polypeptide from sika antler plate and activities of antitumor.
abstract: 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 10,646 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 G0/G1 is 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.
authors.affiliation: College of Life Sciences, Jilin Agriculture University, No. 2888 Xincheng Road, Changchun, Jilin province, 130118, China. weisdhu@163.com.
authors.forename: Wei
authors.initials: W
authors.lastname: Hu
authors.affiliation: College of Life Sciences, Jilin Agriculture University, No. 2888 Xincheng Road, Changchun, Jilin province, 130118, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Qi
authors.affiliation: China Agriculture University, Beijing, 100094, China.
authors.forename: Yu H
authors.initials: YH
authors.lastname: Tian
authors.affiliation: The branch institute of Liaodong college agronomy courtyard, Dandong, 118001, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Hu
authors.affiliation: College of Life Sciences, Jilin Agriculture University, No. 2888 Xincheng Road, Changchun, Jilin province, 130118, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Wu
authors.affiliation: College of Life Sciences, Jilin Agriculture University, No. 2888 Xincheng Road, Changchun, Jilin province, 130118, China.
authors.forename: Xing Y
authors.initials: XY
authors.lastname: Meng
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D000999:Antlers / Q000737:chemistry*; D002453:Cell Cycle / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D003670:Deer; D016193:G1 Phase / Q000187:drug effects; D008297:Male; D015674:Mammary Neoplasms, Animal / Q000188:drug therapy / Q000201:enzymology / Q000473:pathology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*; D051381:Rats; D016192:Resting Phase, Cell Cycle / Q000187:drug effects; D019098:Telomerase / Q000235:genetics / Q000378:metabolism
chemical_list: D000970:Antineoplastic Agents; D010455:Peptides; D019098:Telomerase