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Effect on cancer cells of plasmids that express antisense RNA of human papillomavirus type 18
C Steele1, P G Sacks, K Adler-Storthz
1Department of Microbiology, University of Texas Dental Branch, Houston 77225.
Abstract:
Some human squamous cell carcinomas contain DNA of human papillomaviruses (HPV) and express RNA from the E6 and E7 genes. We have examined the effect of plasmids that express antisense RNA of these genes on the growth of the human cancer cell lines HeLa, C4-1, and 1483, which contain HPV type 18 DNA. As controls, the human cancer cell line 183 and the Vero line of monkey kidney cells were used, which do not contain HPV. Plasmids were introduced into the cells by electroporation; cells that contained HPV type 18 accepted the antisense-expressing plasmids at a lower frequency than the cells that lacked HPV. Cell lines were developed from HeLa cells that contained sense- or antisense-expressing plasmids, and lines that contained antisense-expressing plasmids showed slower growth, reduced ability to form colonies in soft agar, and increased serum requirements. The use of antisense HPV RNA might be a suitable approach to gene therapy of HPV-expressing human cancers.
Insights
Antisense RNA targeting human papillomaviruses (HPV) E6 and E7 genes inhibited growth in HPV-positive cancer cells. This suggests antisense HPV RNA therapy could treat HPV-expressing cancers.
Area of Science:
- Oncology
- Virology
- Molecular Biology
Background:
- Human papillomaviruses (HPV) are linked to squamous cell carcinomas.
- HPV E6 and E7 genes are expressed in some human cancers.
Purpose of the Study:
- To investigate the effect of antisense RNA on HPV-18 DNA-containing cancer cell growth.
- To evaluate antisense HPV RNA as a potential gene therapy for HPV-associated cancers.
Main Methods:
- Electroporation of plasmids expressing sense or antisense RNA for HPV E6/E7 genes into human cancer cell lines (HeLa, C4-1, 1483) and control cells.
- Development of stable cell lines expressing sense or antisense plasmids.
- Assessment of cell growth, colony formation in soft agar, and serum requirements.
Main Results:
- Cells containing HPV type 18 accepted antisense-expressing plasmids less frequently than HPV-negative cells.
- Antisense HPV RNA-expressing cell lines exhibited slower growth.
- Reduced colony formation in soft agar and increased serum requirements were observed in antisense HPV RNA-expressing cells.
Conclusions:
- Antisense HPV RNA expression inhibits the proliferation of HPV-18-positive cancer cells.
- Antisense HPV RNA represents a promising strategy for gene therapy in HPV-expressing human cancers.