|Budget Amount *help
¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1991: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1990: ¥700,000 (Direct Cost: ¥700,000)
In order to investigate the role of oncogenes in the carcinogenesis of skin cancers, certain activatedoncogenes have been investigated by polymerase chain reaction(PCR)method. However, ras genes, which have been most frequently detected in human neoplasmas, were not frequently activated in skin cancers. Instead of investigating oncogenes, human T lymphotropic virus(HTLV-I)and human papillomavirus(HPV)were examined for their involvement in skin cancers, using PCR method.
1. DNAs were extracted from formalin-fixed and paraffin-embedded skin tissues(samples ; 10 lymphoma, 2 verrucous carcinoma, and I bowenoid papulosis).
2. Primers for PCR were chosen for highly conserved regions of HTLV-I and HPV.
3. PCR was performed in a thermal cycler, using the standard method.
4. Amplified DNA fragments were detected in the gels stained by ethidium bromide or by Southern-blot analysis.
1. DNAs were successfully extracted from formalin-fixed and paraffin-embedded tissues, and target genes could be amplified by the PCR.
2. DNA fragments were positively amplified in the tissue s of 2 lymphoma(HTLV-I), 2 verrucous carcinoma(HPV type-16), and I bowenoid papulosis(HPV type-16).
Because the frequency of ras gene activation had been defined to be relatively low in skin cancers, it seemed very difficult to demonstrate the relationship between aging and carcinogenesis through oncocgene activation. In the present investigation, we can not obtain the results of proposed purposes, however, it was demonstrated that the PCR is a very powerful technique to investigate the some particular genes which are even contained in formalin-fixed and paraffin-embedded tissues.