title: Expression, purification and anticancer analysis of GST-tagged human perforin and granzyme B proteins in human laryngeal cancer Hep-2 cells.
abstract: Granzyme B and perforin, two major effector molecules in the granule-mediated cytolytic pathway, are thought to be involved in suppression of tumor progression. In this study, the pGEX-4T-1 expression vector was used to express full-length human perforin or granzyme B as a GST-tagged fusion protein in Escherichia coli (E. coli). GST-tagged proteins were induced with IPTG and purified by GSTrap 4B columns. Purified fusion proteins migrated at the predicted molecular mass on SDS-PAGE and were recognized by specific antibodies. Moreover, the fusion proteins can induce apoptosis and directly inhibit the growth of human laryngeal cancer Hep-2 cells in vitro. These results suggest that active perforin and granzyme B fusion proteins can be produced in E. coli and exhibit anticancer potential in laryngeal cancer cells.
authors.affiliation: Institute of Genomic Medicine, College of Pharmacy, Jinan University, 601 Huangpudadao Xi Road, Guangzhou 510632, China.
authors.forename: Xiuying
authors.initials: X
authors.lastname: Li
authors.affiliation: Institute of Genomic Medicine, College of Pharmacy, Jinan University, 601 Huangpudadao Xi Road, Guangzhou 510632, China.
authors.forename: Guang
authors.initials: G
authors.lastname: Zhang
authors.affiliation: Institute of Genomic Medicine, College of Pharmacy, Jinan University, 601 Huangpudadao Xi Road, Guangzhou 510632, China.
authors.forename: Guijie
authors.initials: G
authors.lastname: An
authors.affiliation: Institute of Genomic Medicine, College of Pharmacy, Jinan University, 601 Huangpudadao Xi Road, Guangzhou 510632, China.
authors.forename: Sha
authors.initials: S
authors.lastname: Liu
authors.affiliation: Institute for Chemical Carcinogenesis, Guangzhou Medical University, 195 Dongfeng Xi Road, Guangzhou 510182, China. Electronic address: lai.yandong@hotmail.com.
authors.forename: Yandong
authors.initials: Y
authors.lastname: Lai
mesh_terms: D000970:Antineoplastic Agents / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004926:Escherichia coli / Q000235:genetics; D005982:Glutathione Transferase; D053804:Granzymes / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007822:Laryngeal Neoplasms; D054353:Perforin / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology*
chemical_list: D000970:Antineoplastic Agents; D011993:Recombinant Fusion Proteins; D054353:Perforin; D005982:Glutathione Transferase; D053804:Granzymes
keywords: Apoptosis; Expression; Granzyme B; Perforin; Purification