Analysis of the human carcinoembryonic antigen promoter core region in colorectal carcinoma-selective cytosine deaminase gene therapy.
We isolated a 204-base pair carcinoembryonic antigen (CEA) promoter core region from a CEA-producing human colorectal carcinoma (CRC) and constructed retrovirus vectors carrying the expression cassette consisting of the CEA promoter core region and the cytosine deaminase (CD) gene. pCD2 retrovirus carrying the CD gene directed by the retrovirus long terminal repeat promoter served as a control vector. An in vitro study showed that the CEA promoter conferred CEA-producing cell-selective CD expression, specifically when the CD expression cassette was inserted into the 3' long terminal repeat of the retrovirus vector. CD-modified CRC xenografts in nude mice were sensitive to 5-fluorocytosine and caused a profound bystander effect on the unmodified CRC. When nude mice harboring intraperitoneally disseminated CRCs were injected intraperitoneally with the CD expression cassette-carrying retrovirus-producing cells, CD transduction into the disseminated CRCs and bone marrow (BM) was observed. CD expression was, however, restricted to CRCs, and it was observed in both CRCs and BM of mice injected with pCD2 retrovirus-producing cells, resulting in better therapeutic outcomes without BM suppression. These results indicate that effective and safe in vivo gene therapy for CRC may be feasible by transferring the CD gene controlled by the CEA promoter core region.
Cao
G
G
Department of Microbiology, Second Military Medical University, Shanghai, China.
Kuriyama
S
S
Cui
L
L
Nagao
S
S
Pan
X
X
Toyokawa
Y
Y
Zhang
X
X
Nishiwaki
I
I
Qi
Z
Z
D000818:Animals; D017209:Apoptosis / Q000235:genetics; D001483:Base Sequence; D001853:Bone Marrow / Q000473:pathology; D002272:Carcinoembryonic Antigen / Q000235:genetics*; D015179:Colorectal Neoplasms / Q000473:pathology / Q000628:therapy*; D043525:Cytosine Deaminase; D017931:DNA Primers; D015316:Genetic Therapy; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D009700:Nucleoside Deaminases / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012190:Retroviridae / Q000235:genetics; D014407:Tumor Cells, Cultured
D002272:Carcinoembryonic Antigen; D017931:DNA Primers; D009700:Nucleoside Deaminases; D043525:Cytosine Deaminase