TY - JOUR
T1 - Adenovirus-mediated transfer of HPV 16 E6/E7 antisense RNA to human cervical cancer cells
AU - Hamada, Katsuyuki
AU - Sakaue, Morito
AU - Alemany, Ramon
AU - Zhang, Wei Wei
AU - Horio, Yoshitsugu
AU - Roth, Jack A.
AU - Mitchell, Michele Follen
N1 - Funding Information:
1This study was partially supported by a grant from the National Cancer Institute (RO1 CA45187) [J.A.R.]; by gifts to the Division of Surgery from Tenneco and Exxon for the Core Laboratory Facility, by the M. D. Anderson Cancer Center Core Support Grant (CA 16672); by grants from the Mathers Foundation, the University Cancer Foundation, and the Business and Professional Women’s Fund; and by a sponsored research agreement with Introgen Therapeutics, Inc.
PY - 1996/11
Y1 - 1996/11
N2 - To explore the potential of an adenoviral antisense RNA transcript for gene therapy of cervical cancer, we introduced the antisense RNA transcript of E6 and E7 genes of human papillomavirus (HPV) 16 into cervical cancer cells harboring HPV 16 via a recombinant adenoviral vector, Ad5CMV-HPV 16 AS and analyzed the effects of expression of these genes on cell growth and tumor growth. Ad5CMV-HPV 16 AS contains the cytomegalovirus-promoter, E6 and E7 genes of HPV 16 in antisense orientation, and the SV40 polyadenylation signal in a mini-gene cassette, which is inserted into the E1-deleted region of modified adenovirus 5. The entire E6/E7 region of HPV 16 was amplified by polymerase chain reaction (PCR) before cloning into the mini-gene cassette. By reverse transcriptase-PCR, HPV 16 E6/E7 antisense RNA was detected in SiHa cells infected with Ad5CMV-HPV 16 AS. The growth of the Ad5CMV-HPV 16 AS- infected cells was greatly suppressed, as evidenced by a decrease in cell count. The growth inhibitory effect of Ad5CMV-HPV 16 AS was significantly enhanced by an adenoviral p53 construct, Ad5CMV-p53. In an ex vivo study in nude mice, tumorigenicity was completely inhibited in mice injected with Ad5CMV-HPV 16 AS-infected SiHa cells. These data suggest that transfection of cervical cancer cells with HPV 16 E6/E7 antisense RNA in a form such as Ad5CMV-HPV 16 AS is a potential novel approach to the therapy of HPV 16- positive cervical cancer.
AB - To explore the potential of an adenoviral antisense RNA transcript for gene therapy of cervical cancer, we introduced the antisense RNA transcript of E6 and E7 genes of human papillomavirus (HPV) 16 into cervical cancer cells harboring HPV 16 via a recombinant adenoviral vector, Ad5CMV-HPV 16 AS and analyzed the effects of expression of these genes on cell growth and tumor growth. Ad5CMV-HPV 16 AS contains the cytomegalovirus-promoter, E6 and E7 genes of HPV 16 in antisense orientation, and the SV40 polyadenylation signal in a mini-gene cassette, which is inserted into the E1-deleted region of modified adenovirus 5. The entire E6/E7 region of HPV 16 was amplified by polymerase chain reaction (PCR) before cloning into the mini-gene cassette. By reverse transcriptase-PCR, HPV 16 E6/E7 antisense RNA was detected in SiHa cells infected with Ad5CMV-HPV 16 AS. The growth of the Ad5CMV-HPV 16 AS- infected cells was greatly suppressed, as evidenced by a decrease in cell count. The growth inhibitory effect of Ad5CMV-HPV 16 AS was significantly enhanced by an adenoviral p53 construct, Ad5CMV-p53. In an ex vivo study in nude mice, tumorigenicity was completely inhibited in mice injected with Ad5CMV-HPV 16 AS-infected SiHa cells. These data suggest that transfection of cervical cancer cells with HPV 16 E6/E7 antisense RNA in a form such as Ad5CMV-HPV 16 AS is a potential novel approach to the therapy of HPV 16- positive cervical cancer.
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U2 - 10.1006/gyno.1996.0310
DO - 10.1006/gyno.1996.0310
M3 - Article
C2 - 8910631
AN - SCOPUS:0030298504
SN - 0090-8258
VL - 63
SP - 219
EP - 227
JO - Gynecologic oncology
JF - Gynecologic oncology
IS - 2
ER -