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[Molecular cloning and expression of anti-tumor adhesion peptide (beta3)].

Abstract
To block tumor cell adhesion, inhibit tumor metastasis and recurrence, the anti-adhesion peptide-trimeric beta peptide (DLYYLMDLSYSMKGGDLYYLMDLSYSMKGGDLYYLMDLSYSMK, beta3) was designed. The DNA fragment of beta3 was cloned into expression vector pET-His and the fusion protein His-beta3 was expressed in E. coli. BL21(DE3)plysS. After 1.5 hours' induction with IPTG, His-beta3 peptide was expressed significantly amounting to 10% of the insoluble proteins and 4% of the total proteins. 20mg of beta3 peptide was obtained from one litter culture medium after purification by using metal-chelating sepharose 6B FF. The purity of beta3 is 92.2% according to Gel-Pro analysis. The anti-adhesion effects of beta3 peptide, beta1 peptide (DLYYLMDLSYSMK) and GRGDS on the hepatocellular carcinoma cell line SMMC-7721 and the high metastasis hepatocellular carcinoma cell line HCCLM6 were studied. The result showed the beta3 blocked the adhesion of HCCLM6 cells and SMMC-7721 cells to fibronectin (FN) specifically. The inhibition effect was dose-dependent and time-dependent and the inhibition rate of beta3 was higher than three times concentration of beta1 and GRGDS. This suggested that pET-His-beta3/BL21(DE3)plysS was a suitable expression system for beta3, and the expressed beta3 specially inhibited the adhesion of cancer cells.
AuthorsSong-Mei Wang, Jun Zhu, Yan Li, Luan-Feng Pan, Xi-Liang Zha, Yin-Kun Liu
JournalSheng wu gong cheng xue bao = Chinese journal of biotechnology (Sheng Wu Gong Cheng Xue Bao) Vol. 21 Issue 4 Pg. 558-62 (Jul 2005) ISSN: 1000-3061 [Print] China
PMID16176092 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Antineoplastic Agents
  • Peptide Fragments
  • Peptides
  • Recombinant Proteins
Topics
  • Amino Acid Sequence
  • Antineoplastic Agents (pharmacology)
  • Cell Adhesion (drug effects)
  • Cloning, Molecular
  • Escherichia coli (genetics, metabolism)
  • Molecular Sequence Data
  • Peptide Fragments (pharmacology)
  • Peptides (genetics, metabolism, pharmacology)
  • Recombinant Proteins (biosynthesis, genetics)
  • Tumor Cells, Cultured

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