title: Potential antitumor therapeutic application of Grimontia hollisae thermostable direct hemolysin mutants.
abstract: We report on the preparation of a new type of immunotoxin by conjugation of an epidermal growth factor receptor (EGFR)-binding peptide and an R46E mutation of thermostable direct hemolysin from Grimontia hollisae, (Gh-TDH(R) (46E) /EB). The hybrid immunotoxin was purified to homogeneity and showed a single band with slight slower mobility than that of Gh-TDH(R) (46E) . Cytotoxicity assay of Gh-TDH(R) (46E) /EB on EGFR highly, moderately, low, and non-expressed cells, A431, MDA-MB-231, HeLa, and HEK293 cells, respectively, showed apparent cytotoxicity on A431 and MDA-MB-231 cells but not on HeLa or HEK293 cells. In contrast, no cytotoxicity was observed for these cells treated with either Gh-TDH(R) (46E) or EB alone, indicating enhanced cytotoxic efficacy of Gh-TDH(R) (46E) by the EGFR binding moiety. Further antitumor activity assay of Gh-TDH(R) (46E) /EB in a xenograft model of athymic nude mice showed obvious shrinkage of tumor size and degeneration, necrosis, and lesions of tumor tissues compared to the normal tissues. Therefore, the combination of Gh-TDH(R) (46E) with target affinity agents opens new possibilities for pharmacological treatment of cancers and potentiates the anticancer drug's effect.
authors: Department of Biological Science and Technology, National Chiao Tung University, Hsin-Chu, Taiwan.
authors: Sheng-Cih
authors: SC
authors: Huang
authors: Yu-Kuo
authors: YK
authors: Wang
authors: Wan-Ting
authors: WT
authors: Huang
authors: Tsam-Ming
authors: TM
authors: Kuo
authors: Bak-Sau
authors: BS
authors: Yip
authors: Tien-Hsiung Thomas
authors: TH
authors: Li
authors: Tung-Kung
authors: TK
authors: Wu
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics / Q000494:pharmacology*; D001427:Bacterial Toxins / Q000235:genetics / Q000494:pharmacology; D045744:Cell Line, Tumor; D066246:ErbB Receptors / Q000235:genetics* / Q000378:metabolism; D005260:Female; D057809:HEK293 Cells; D006367:HeLa Cells; D006460:Hemolysin Proteins / Q000235:genetics / Q000494:pharmacology*; D006801:Humans; D000922:Immunotoxins / Q000494:pharmacology*; D051379:Mice; D008819:Mice, Nude; D011994:Recombinant Proteins / Q000235:genetics / Q000494:pharmacology*; D014737:Vibrionaceae / Q000235:genetics
chemical_list: D000970:Antineoplastic Agents; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D006460:Hemolysin Proteins; D000922:Immunotoxins; D011994:Recombinant Proteins; C073845:hemolysin, Vibrio; C068489:thermostable direct hemolysin; D066246:ErbB Receptors
keywords: Anticancer; Grimontia hollisae; epidermal growth factor receptor; immunotoxin; thermostable direct hemolysin
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