Oligopeptide with breast cancer resistant activity and application thereof
An anti-breast cancer and oligopeptide technology, applied in the field of biomedicine, can solve the problems of high cost and long cycle, and achieve the effects of small molecular weight, inhibition of proliferation, and easy industrialization
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Embodiment 1
[0024] Design and synthesis of oligopeptides
[0025] 1. Extract the main international antimicrobial peptide database (http: / / aps.unmc.edu / AP / main.php; http: / / penbase.immunaqua.com / ; http: / / www.bbcm.univ.trieste.it / ; http: / / www.imtech.res.in / raghava / antibp / ) registered in the antibacterial peptide sequence, analyze the corresponding relationship between its primary structure and activity, and design artificial short peptide sequence.
[0026] . Utilize the protein analysis database http: / / www.expasy.org / to analyze the similarity between the oligopeptide sequence with known activity and the secondary structure of the designed oligopeptide sequence, and correct the designed sequence in combination with other physicochemical parameter predictions.
[0027] , The oligopeptide sequence is sent to the biological company for synthesis or the full sequence is synthesized by an automatic oligopeptide synthesizer. The purity of the HPLC test should be greater than 96%, and t...
Embodiment 2
[0031] Identification of oligopeptides
[0032] P1221 oligopeptide was hydrolyzed with 5.7mol / L HCl for 16 hours, and amino acid analysis was carried out. It was known that it contained 9 kinds of amino acids, including His, Asn, Tyr, Gly, Phe, Val, Pro, Ser, and Leu, and the molar ratio was His:Asn :Tyr:Gly:Phe:Val:Pro:Ser:Leu = 1:1:1:1:1:1:1:1:1. After amino acid sequence analysis, the sequence is His-Asn-Tyr-Gly-Phe-Val-Pro-Ser-Leu. A peak was obtained by electrospray mass spectrometry, and its molecular weight was 1033.15.
Embodiment 3
[0034] Detection of tumor suppressor activity of P1221 oligopeptide in vitro by MTT assay
[0035] 2.1 Materials and reagents
[0036] 1) Oligopeptide P1221 was synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd.
[0037] ) Human breast cancer cell lines MCF-7, MDA-MB-231, and human normal breast cell lines MCF-10A were purchased from the Cell Resource Center, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences.
[0038] ) RPMI 1640 medium, DMEM medium, and newborn calf serum are all products of American GIBCO Company.
[0039] ) dimethyl sulfoxide (DMSO), tetramethylazozole (MTT) and trypsin are products of Sigma.
[0040] 2.2 Experiment
[0041] 1) Dilute the P1221 oligopeptide to an initial concentration of 1 mM with a diluent (sterile PBS, or sterile water, or the corresponding cell culture medium), filter and sterilize, and freeze at -20°C after aliquoting.
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