Growth inhibition of human cervical cancer cells with the recombinant adenovirus p53 in vitro.
Human papillomavirus (HPV) has been identified in the majority of invasive cancers of the uterine cervix sampled and has been found to contribute in a significant way to the genesis of human cervical cancer. HPV has two transforming genes that encode the oncoproteins E6 and E7. E6 can form complexes with p53 and promote p53 degradation. We introduced wild-type p53 into a cervical cancer cell line via a recombinant adenoviral vector, Ad5CMV-p53. Human cervical cancer cell line HeLa, which has HPV type 18 and wild-type p53, was used in this study. Cells were grown in RPMI medium supplemented with 10% heat-inactivated fetal bovine serum. Ad5CMV-p53 was created by inserting the cytomegalovirus promoter, wild-type p53 cDNA, and SV40 polyadenylation signal in a minigene cassette into the E1-deleted region of the modified Ad5 adenovirus. The transduction efficiency was 100% when a dose ensuring a multiplicity of infection of 100 or greater was used. The p53 protein was detected in Ad5CMV-p53-infected cells by immunohistochemical and Western blot analyses. The growth of the Ad5CMV-p53-infected cells was greatly suppressed as detected by both cell count and [3H]thymidine incorporation assay. These data suggest that transfection of HPV-positive cervical cancer cells with a wild-type p53 gene in a form such as Ad5CMV-p53 is a potential novel therapy for cervical cancer.
Hamada
K
K
Department of Thoracic and Cardiovascular Surgery, University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.
Zhang
W W
WW
Alemany
R
R
Wolf
J
J
Roth
J A
JA
Mitchell
M F
MF
D000256:Adenoviridae / Q000235:genetics*; D002277:Carcinoma / Q000235:genetics / Q000473:pathology*; D002452:Cell Count; D002455:Cell Division; D005260:Female; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D011994:Recombinant Proteins; D016159:Tumor Suppressor Protein p53 / Q000235:genetics* / Q000378:metabolism*; D002583:Uterine Cervical Neoplasms / Q000235:genetics / Q000473:pathology*
D011994:Recombinant Proteins; D016159:Tumor Suppressor Protein p53