Molecular cloning and expression of anti-tumor adhesion peptide (beta3) / 生物工程学报
Chinese Journal of Biotechnology
; (12): 558-562, 2005.
Article
in Zh
| WPRIM
| ID: wpr-305203
Responsible library:
WPRO
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.
Full text:
1
Index:
WPRIM
Main subject:
Peptide Fragments
/
Peptides
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Pharmacology
/
Recombinant Proteins
/
Molecular Sequence Data
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Tumor Cells, Cultured
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Cell Adhesion
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Amino Acid Sequence
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Cloning, Molecular
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Escherichia coli
Language:
Zh
Journal:
Chinese Journal of Biotechnology
Year:
2005
Type:
Article