Site-directed mutagenesis of human IL-29 and antineoplastic activity of the recombinant human IL-29 variant.
- Author:
Wei CHEN
;
Rong ZHU
;
Chunlei GE
;
Yuan LU
;
Liyun LI
;
Fei LI
;
Minchen WU
- Publication Type:Journal Article
- MeSH:
Antineoplastic Agents;
pharmacology;
Carcinoma, Hepatocellular;
pathology;
Cell Line, Tumor;
drug effects;
Humans;
Interleukins;
biosynthesis;
pharmacology;
Liver Neoplasms;
pathology;
Mutagenesis, Site-Directed;
Pichia;
Plasmids;
Polymerase Chain Reaction;
Recombinant Proteins;
biosynthesis;
pharmacology
- From:
Chinese Journal of Biotechnology
2015;31(5):702-710
- CountryChina
- Language:Chinese
-
Abstract:
To explore the anti-tumor proliferation activity of human interleukin-29 (hIL-29) variant and based on bioinformatics analyzed data of hIL-29, a mutant gene hIL-29(mut33,35) was amplified by site-directed mutagenesis and megaprimer PCR. The hIL-29(mut33,35) was inserted into an eukaryotic expression plasmid pPIC9K and successfully expressed in Pichia pastoris GS115. A recombinant variant protein (rhIL-29(mut33,35)) was purified from the ferment supernatant of the engineering GS115. To observe the antineoplastic activity of the variant rhIL-29(mut33,35), a CCK-8 reagent was used to detect the anti-proliferation effect. Results show that it has strong anti-proliferation effect when acted on liver cancer cell BEL7402, colon cancer cell HCT8 and gastric cancer cell SGC7901. The inhibition ratios of the three tumor cells were (30.99 ± 1.58)%, (22.47 ± 1.37)% and (32.05 ± 2.02)%, respectively. In high dose group, the anti-proliferation effect of the rhIL-29(mut33,35) was stronger than that of wild type rhIL-29 (P < 0.01). This indicates the variant rhIL-29(mut33,35) has potential development value for medicine.
