title: Expression and purification of the recombinant porcine NK-lysin in Pichia pastoris and observation of anticancer activity in vitro.
abstract: Natural killer (NK)-lysin, a broad-spectrum antimicrobial peptide, has antitumor and antibactericidal activities against both gram-positive and gram-negative bacteria. In this study the recombinant porcine NK-lysin was expressed and purified in the Pichia pastoris system, and then 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide was used to assess its anticancer activity in vitro. The results showed that the recombinant porcine NK-lysin possesses potent antitumor activity against the human hepatocellular carcinoma cell line SMMC-7721 in a time- and dose-dependent manner, but has negligible hemolysis activity against human erythrocytes. Scanning electronic microscopy was used to directly observe the ultrastructure of SMMC-7721 cells treated with NK-lysin; untreated cells showed lamellipodia and filopodia scattered with the cell surface, with good cell-cell contacts among neighboring cells. In contrast, treated tumor cells exhibited marked alterations in cell morphology, and cell-cell contacts disappeared among neighboring cells. Compared with the untreated tumor cells, the tumor cells treated with NK-lysin for 12 and 24 hr were suppressed for the expression of fascin 1. Thus, the recombinant porcine NK-lysin potentially could be developed as a therapeutic agent for inhibiting tumor growth.
authors.affiliation: a College of Animal Science and Veterinary Medicine , Shanxi Agricultural University , Taigu , Shanxi , China.
authors.forename: Kuohai
authors.initials: K
authors.lastname: Fan
authors.affiliation: a College of Animal Science and Veterinary Medicine , Shanxi Agricultural University , Taigu , Shanxi , China.
authors.forename: Hongquan
authors.initials: H
authors.lastname: Li
authors.affiliation: b Transplantation Biology Research Center , Massachusetts General Hospital and Harvard Medical School , Boston , Massachusetts , USA.|c MGH-DF/HCC Recombinant Protein Expression and Purification Core , Boston , Massachusetts , USA.
authors.forename: Zhirui
authors.initials: Z
authors.lastname: Wang
authors.affiliation: a College of Animal Science and Veterinary Medicine , Shanxi Agricultural University , Taigu , Shanxi , China.
authors.forename: Wenjuan
authors.initials: W
authors.lastname: Du
authors.affiliation: a College of Animal Science and Veterinary Medicine , Shanxi Agricultural University , Taigu , Shanxi , China.
authors.forename: Wei
authors.initials: W
authors.lastname: Yin
authors.affiliation: a College of Animal Science and Veterinary Medicine , Shanxi Agricultural University , Taigu , Shanxi , China.
authors.forename: Yaogui
authors.initials: Y
authors.lastname: Sun
authors.affiliation: a College of Animal Science and Veterinary Medicine , Shanxi Agricultural University , Taigu , Shanxi , China.
authors.forename: Junbing
authors.initials: J
authors.lastname: Jiang
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D045744:Cell Line, Tumor; D006801:Humans; D066298:In Vitro Techniques; D010843:Pichia / Q000235:genetics*; D011510:Proteolipids / Q000235:genetics* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000235:genetics / Q000494:pharmacology; D013552:Swine
chemical_list: D000970:Antineoplastic Agents; C093238:NK-lysin; D011510:Proteolipids; D011994:Recombinant Proteins
keywords: Antitumor; Pichia pastoris; expression; fascin 1; porcine NK-lysin; purification