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.
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Authors | Song-Mei Wang, Jun Zhu, Yan Li, Luan-Feng Pan, Xi-Liang Zha, Yin-Kun Liu |
Journal | Sheng 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 |
PMID | 16176092
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Antineoplastic Agents
- Peptide Fragments
- Peptides
- Recombinant Proteins
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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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