AIM:To establish a tissue-specific gene therapy for colorectal carcinoma using bacterial ADP-ribosylating toxin genes. METHODS: Pseudomonas exotoxin A domain [II+III (PEA) was cloned from genomic DNA of Pseudomonas aeruginosa.PEA and diphtheria toxin A chain gene (DTA) were modified to express eukaryotically. After sequencing, the toxin genes under the control of human carcinoembryonic antigen (CEA)promoter were cloned into retroviral vectors to construct CEAPEA and CEADTA respectively. In vitro cotransfection of the constructs with luciferase vectors and in vivo gene transfer in nude mice were subsequently carried out. RESULTS: Both CEAPEA and CEADTA specifically inhibited the reporter gene expression in the CEA positive human colorectal carcinoma (CRC) cells in vitro. Direct injection of CEAPEA and CEADTA constructs into the established human tumors in BALB/c nude mice led to significant and selective reductions in CRC tumor size as compared with that in control groups. CONCLUSION: The toxin genes, working as therapeutic genes, are suitable for the tissue-specific gene therapy for colorectal carcinoma.
AIM:To establish a tissue-specific gene therapy for colorectal carcinoma using bacterial ADP-ribosylating toxin genes. METHODS:Pseudomonas exotoxin A domain [II+III (PEA) was cloned from genomic DNA of Pseudomonas aeruginosa.PEA and diphtheria toxin A chain gene (DTA) were modified to express eukaryotically. After sequencing, the toxin genes under the control of human carcinoembryonic antigen (CEA)promoter were cloned into retroviral vectors to construct CEAPEA and CEADTA respectively. In vitro cotransfection of the constructs with luciferase vectors and in vivo gene transfer in nude mice were subsequently carried out. RESULTS: Both CEAPEA and CEADTA specifically inhibited the reporter gene expression in the CEA positive humancolorectal carcinoma (CRC) cells in vitro. Direct injection of CEAPEA and CEADTA constructs into the established humantumors in BALB/c nude mice led to significant and selective reductions in CRC tumor size as compared with that in control groups. CONCLUSION: The toxin genes, working as therapeutic genes, are suitable for the tissue-specific gene therapy for colorectal carcinoma.