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dc.contributor.authorYao, Qin
dc.contributor.authorCao, Siyu
dc.contributor.authorLi, Chun
dc.contributor.authorKong, Beihua
dc.contributor.authorDai, Shuzhen
dc.contributor.authorWei, Mingqian
dc.date.accessioned2017-05-03T13:50:00Z
dc.date.available2017-05-03T13:50:00Z
dc.date.issued2009
dc.date.modified2010-07-09T03:06:09Z
dc.identifier.issn1099-498X
dc.identifier.urihttp://hdl.handle.net/10072/31629
dc.description.abstractClaudin-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.publicationstatusYes
dc.languageEnglish
dc.language.isoeng
dc.publisherJohn Wiley & Sons, Inc.
dc.publisher.placeUnited States
dc.relation.ispartofstudentpublicationN
dc.relation.ispartofconferencename6th Meeting of the Australasian-Gene-Therapy-Society
dc.relation.ispartofconferencetitleJOURNAL OF GENE MEDICINE
dc.relation.ispartofdatefrom2009-04-29
dc.relation.ispartofdateto2009-05-01
dc.relation.ispartoflocationSydney, AUSTRALIA
dc.relation.ispartofpagefrom853
dc.relation.ispartofpagefrom1 pages
dc.relation.ispartofpageto853
dc.relation.ispartofpageto1 pages
dc.relation.ispartofissue9
dc.relation.ispartofvolume11
dc.rights.retentionY
dc.subject.fieldofresearchClinical sciences
dc.subject.fieldofresearchClinical sciences not elsewhere classified
dc.subject.fieldofresearchcode3202
dc.subject.fieldofresearchcode320299
dc.titleRecombinant Cytotoxin C-CPE-ETA' Effectively Kills Claudin-4-Expressing Cancer Cells
dc.typeConference output
dc.type.descriptionE3 - Conferences (Extract Paper)
dc.type.codeE - Conference Publications
gro.facultyGriffith Health, School of Medical Science
gro.date.issued2009
gro.hasfulltextNo Full Text
gro.griffith.authorWei, Ming Q.


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