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Nucleic Acids Research, 1991, Vol. 19, No. 15 4109-4114
© 1991

MOLECULAR BIOLOGY

Anti-sense phosphorothioate oligonucleotides have both specific and non-specific effects on cells containing human papillomavirus type 16

Alan Storey, David Oates1, Lawrence Banks, Lionel Crawford1 and Tim Crook2

International Center for Genetic Engineering and Biotechnology Padriciano 99, I-34012 Trieste, Italy 1ICRF Tumour Virus Group, Department of Pathology, Universfty of Cambridge Tennis Court Road, Cambridge CB2 1QP UK 2Ludwig Institute for Cancer Research, St Mary's Hospital Medical School Norfolk Place, London W2 1PG, UK

Received May 23, 1991. Accepted July 2, 1991.

A range of specific nuclease resistant phosphorothioate oligodeoxynucleotides (S-oligos) complementary to mRNA of human papillomavirus type 16 (HPV16), were tested for their ability to inhibit cell proliferation and to alter the level of HPV-specific mRNA and proteins in CaSki cells, a human cervical carcinoma cell line containing HPV16 DNA. Only certain of the S-oligos to the viral upstream regulatory region (URR) and the early viral open reading frames (ORF), E6 and E7, were found to display any activity on the ceiis. These S-oligos were found to exhibit potent anti proliferative activity at concentrations between 0.25DµM and 20µM inhibiting the uptake of [3H]-thymidine into CaSki cells by up to 90% at higher concentrations. The rate of synthesis of E6 and E7 proteins and the steady state level of the E7 protein however remained largely unchanged. E7 protein exhibited a greater decrease in phosphorylation in the presence of only one of the antisense oligos. Other S-oligos including a random sequence, unmodified sequences or O-methylphosphonate modified oligos, had no specific effect on the cells. The results impiy that the anti-sense S-oligo nucleotides had both specific anti-HPV16 and other non-specific effects on cell proliferation and synthesis of viraily encoded proteins.


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