A Study on Construction of RGD Motif in Human Interleukin-18 and Its Inhibitory Activity on B16 Melanoma
- VernacularTitle:人白介素18(IL-18)RGD模体抗黑色素瘤细胞B16活性研究
- Author:
Zhong-Min LU
;
Bao-Chang ZHAO
;
- Publication Type:Journal Article
- Keywords:
Interleukin 18 RGD motif Pichia pastoris Suppression of B16
- From:
China Biotechnology
2006;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Using site-directed mutagenesis and DNA recombinant technology, GGC fragment was inserted into human IL-18 cDNA. The mutated IL-18 cDNA was constructed into plasmid pPIC9K, then transformed into Pichia pastoris GS115 and efficiently expressed. A Glycine residue was inserted into the recombinant IL-18 between Arg39 and Asp40 to form a RGD motif. The mutated IL-18 was termed IL-18-RGD. The protein was purified with Sephadex G-100. The cell culture of melanoma B16 showed that IL-18-RGD efficiently inhibited B16 tumor growth, IC50 = 8.10?mol/L, but the inhibitory effect of IL-18 was not detected. Both IL-18-RGD and IL-18 showed inhibitory activities on mice loaded B16 in vivo, the inhibitory efffct of IL-18-RGD was stronger than that of IL-18. The inhibitory activities of IL-18-RGD and IL-18 on bFGF induced angiogenesis on chorioallantoic membranes were detected. There was no differences between IL-18-RGD and wild IL-18 in the activity of inducing IFN-? in human PBMC.
