PMID- 8995542
OWN - NLM
STAT- MEDLINE
DCOM- 19970210
LR  - 20101118
IS  - 0090-8258 (Print)
IS  - 0090-8258 (Linking)
VI  - 64
IP  - 1
DP  - 1997 Jan
TI  - In vitro antigene therapy targeting HPV-16 E6 and E7 in cervical carcinoma.
PG  - 18-25
AB  - Human papillomavirus (HPV) infection is believed to play a central role in 
      cervical carcinogenesis. Specifically, two viral oncoproteins, E6 and E7, possess 
      transforming ability and have been shown to interact with the cellular tumor 
      suppressors p53 and p105, the retinoblastoma (Rb) gene product. To test the 
      hypothesis that E6 and E7 play an active role in the maintenance of the malignant 
      phenotype and may be ideal targets for antigene therapy, we tested the 
      antiproliferative effects of phosphorothioate oligodeoxynucleotides (oligos) 
      targeting HPV-16 E6 and E7 in cervical cancer cell lines and primary tumor 
      explants. The ATP cell viability assay was used to measure growth effects of 
      27-mer antisense oligos targeting the ATG translational start region of HPV-16 E6 
      and E7 sequences in HPV-16-positive cell lines SiHa and CaSki and four advanced, 
      primary cervical tumor explants. A random oligo sequence, an HPV-18-positive and 
      HPV-negative cell line, one histologically confirmed endometrial and two ovarian 
      tumors were used as negative controls. HPV type was confirmed by hybrid capture 
      techniques. Cell lines and sterile (staging laparotomy) tumor cells were plated 
      at 5000 cells/0.1 ml and 100,000 cells/0.5 ml in 96-well plates or soft agar, 
      respectively, and incubated at 37 degrees C with a single treatment of oligos at 
      0-16 microM. E6/E7 combinations at a fixed ratio of 1:1 were used at 0-8 microM 
      for each oligo. Cellular ATP was measured by luciferin/luciferase fluorescence on 
      Day 6. HPV-16 E6 and E7 oligos showed antiproliferative effects in all 
      HPV-16-positive cell lines and primary tumor explants (IC50s 6.9-9.5 microM for 
      cell lines, 9.1-12.1 microM primary cervical tumors), while the HPV-negative 
      C33-A cell line and HPV-18-positive cell line HeLa were relatively insensitive to 
      the HPV-16 oligos (IC50s > 30 microM extrapolated). The endometrial and two 
      ovarian primary tumors were also insensitive to the HPV E6 and E7 oligos (IC50s > 
      25 microM extrapolated). Random oligos had little effect on cell growth at 
      concentrations up to 16 microM (< 25% inhibition), except in CaSki (@50% 
      inhibition at 16 microM). Combinations of E6 and E7 demonstrated mixed 
      synergistic and antagonistic effects as determined by combination indices (CI) 
      derived from median effect parameters. In the HPV-16-positive primary cervical 
      tumors and the cell line SiHa, E6/E7 combinations were synergistic at low doses 
      (< 25% growth inhibitory dose range) and antagonistic at doses above this. For 
      the HPV-16-positive cell line CaSki, however, E6/E7 combinations were 
      antagonistic at all dose ranges. Phosphorothioate oligos directed against the 
      viral oncogenes E6 and E7 were shown to have antiproliferative effects specific 
      to HPV-containing cancer cells. These specific antiproliferative effects suggest 
      that HPV-16 E6 and E7 sequences play an active role in the malignant growth 
      properties of cervical cancer cells and may be ideal targets for antigene 
      therapy.
FAU - Madrigal, M
AU  - Madrigal M
AD  - Department of Gynecology and Obstetrics, University of Miami School of Medicine, 
      Florida 33136, USA.
FAU - Janicek, M F
AU  - Janicek MF
FAU - Sevin, B U
AU  - Sevin BU
FAU - Perras, J
AU  - Perras J
FAU - Estape, R
AU  - Estape R
FAU - Penalver, M
AU  - Penalver M
FAU - Averette, H E
AU  - Averette HE
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Gynecol Oncol
JT  - Gynecologic oncology
JID - 0365304
RN  - 0 (Antigens, Viral)
RN  - 0 (E6 protein, Human papillomavirus type 16)
RN  - 0 (Oligonucleotides, Antisense)
RN  - 0 (Oncogene Proteins, Viral)
RN  - 0 (Papillomavirus E7 Proteins)
RN  - 0 (Repressor Proteins)
RN  - 0 (Thionucleotides)
RN  - 0 (oncogene protein E7, Human papillomavirus type 16)
SB  - IM
MH  - Antigens, Viral/*therapeutic use
MH  - Dose-Response Relationship, Drug
MH  - Female
MH  - Humans
MH  - Oligonucleotides, Antisense/*therapeutic use
MH  - Oncogene Proteins, Viral/genetics
MH  - Papillomaviridae/genetics/*immunology
MH  - Papillomavirus E7 Proteins
MH  - *Repressor Proteins
MH  - Thionucleotides/*therapeutic use
MH  - Tumor Cells, Cultured
MH  - Uterine Cervical Neoplasms/*therapy/*virology
EDAT- 1997/01/01 00:00
MHDA- 1997/01/01 00:01
CRDT- 1997/01/01 00:00
PHST- 1997/01/01 00:00 [pubmed]
PHST- 1997/01/01 00:01 [medline]
PHST- 1997/01/01 00:00 [entrez]
AID - S0090-8258(96)94515-1 [pii]
AID - 10.1006/gyno.1996.4515 [doi]
PST - ppublish
SO  - Gynecol Oncol. 1997 Jan;64(1):18-25. doi: 10.1006/gyno.1996.4515.