dc.contributor.author | Yao, Qin | |
dc.contributor.author | Cao, Siyu | |
dc.contributor.author | Li, Chun | |
dc.contributor.author | Kong, Beihua | |
dc.contributor.author | Dai, Shuzhen | |
dc.contributor.author | Wei, Mingqian | |
dc.date.accessioned | 2017-05-03T13:50:00Z | |
dc.date.available | 2017-05-03T13:50:00Z | |
dc.date.issued | 2009 | |
dc.date.modified | 2010-07-09T03:06:09Z | |
dc.identifier.issn | 1099-498X | |
dc.identifier.uri | http://hdl.handle.net/10072/31629 | |
dc.description.abstract | Claudin-4 is a natural receptor for Clostridium perfringens enterotoxin (CPE). It has recently been found to be overexpressed on the plasma membrane of a range of cancer cells, such as ovarian, breast, colon and prostate cancer. Hence, we hypothesised that a CPE-based cytotoxin may be of potential use in targeted cytolysis of claudin-4 positive cancers. We created such a novel toxin, C-CPEETA', by fusing C-terminal high affinity binding domain of CPE (C-CPE) with a truncated form of Pseudomonas aeruginosa exotoxin A (ETA'), aiming to examine whether C-CPE will specifically target claudin-4 molecule and the targeted toxin is cytotoxic. After cloning into an expression plasmid and transforming it into a bacterial strain, a 58 kDa recombinant protein was evident by Western blot and Coomassie blue staining. Flow cytometric analysis and immune assays were performed using recombinant C-CPEETA' in a range of claudin-4 positive and negative cancer cell lines. The proapoptotic and cytolytic activities were evaluated using JC-1 staining and MTT assay. Purified and recombinant C-CPE-ETA' bound with high affinity to claudin- 4-positive cancer cells, including breast cancer MCF-7, colon cancer SW480, HT29, prostate cancer DU145, PC3 and ovarian cancer CAOV3. Interestingly, after initial binding, C-CPE-ETA' was sequentially internalized into claudin-4- positive cells within 60 mins, which resulted in significant apoptotic cell death. This apoptotic effects increased when the incubation was further extended to 24h. The IC50 of recombinant C-CPE-ETA' ranged from 0.7 ng/ml to 50 ng/ml in claudin-4-positive cancer cells. However, control claudin-4-negative cell lines, Hela and HUVEC, were not susceptible to the recombinant C-CPE-ETA' at concentrations up to 10姯ml, suggesting the strict claudin-4 selectivity. Further study in a MCF-7 subcutaneous xerograft tumour model system showed that intratumoural administration of recombinant C-CPE-ETA' significantly inhibited tumour growth. Our results suggest that the C-CPE-ETA' exhibits remarkably specific cytotoxicity. Its therapeutic potentials for claudin-4-postitive cancer warrants further development. | |
dc.description.publicationstatus | Yes | |
dc.language | English | |
dc.language.iso | eng | |
dc.publisher | John Wiley & Sons, Inc. | |
dc.publisher.place | United States | |
dc.relation.ispartofstudentpublication | N | |
dc.relation.ispartofconferencename | 6th Meeting of the Australasian-Gene-Therapy-Society | |
dc.relation.ispartofconferencetitle | JOURNAL OF GENE MEDICINE | |
dc.relation.ispartofdatefrom | 2009-04-29 | |
dc.relation.ispartofdateto | 2009-05-01 | |
dc.relation.ispartoflocation | Sydney, AUSTRALIA | |
dc.relation.ispartofpagefrom | 853 | |
dc.relation.ispartofpagefrom | 1 pages | |
dc.relation.ispartofpageto | 853 | |
dc.relation.ispartofpageto | 1 pages | |
dc.relation.ispartofissue | 9 | |
dc.relation.ispartofvolume | 11 | |
dc.rights.retention | Y | |
dc.subject.fieldofresearch | Clinical sciences | |
dc.subject.fieldofresearch | Clinical sciences not elsewhere classified | |
dc.subject.fieldofresearchcode | 3202 | |
dc.subject.fieldofresearchcode | 320299 | |
dc.title | Recombinant Cytotoxin C-CPE-ETA' Effectively Kills Claudin-4-Expressing Cancer Cells | |
dc.type | Conference output | |
dc.type.description | E3 - Conferences (Extract Paper) | |
dc.type.code | E - Conference Publications | |
gro.faculty | Griffith Health, School of Medical Science | |
gro.date.issued | 2009 | |
gro.hasfulltext | No Full Text | |
gro.griffith.author | Wei, Ming Q. | |