H Jin1, J Wang, Y Zuo. 1. Department of Dermatology, Chinese Academy of Medical Sciences, Beijing 100730, China.
Abstract
OBJECTIVE: To study the expression of c-jun, c-fos and MDM2 mRNA in cultured kerotinocytes through transfecting HPV16. METHODS: Keratinocytes were cultured in serum-free medium, and secondary cultures continued in a suitable time. Plasmid pSV(2)-neo/HPV16 was transfected into cultured kerotinocytes using FuGENE(TM)6. Reverse-transcriptase polymerase chain reaction (RT-PCR) was used to detect c-jun, c-fos and MDM2 mRNA in keratinocytes of normal and transfected keratinocytes. RESULTS: After transfection, the morphology and structure of the keratinocytes were changed. The cells became larger and their nuclei became looser. There were particles in the cytoplasm. The expression of HPV16 mRNA was detected in the keratinocytes after transfecting pSV(2)-neo/HPV16 and the product was 110 bp fragment. The DNA fragment (7.9 kb) of HPV16 was also detected. In the transfected keratinocytes, the exprssions of c-jun, c-fos and MDM2 mRNA were detected and the products were 196 bp, 332 bp and 548 bp fragments, respectively. However, the above-mentioned expressions those were not detected in normal keratinocytes. CONCLUSION: HPV leads to cell proliferation by expressing c-jun, c-fos and MDM2.
OBJECTIVE: To study the expression of c-jun, c-fos and MDM2 mRNA in cultured kerotinocytes through transfecting HPV16. METHODS: Keratinocytes were cultured in serum-free medium, and secondary cultures continued in a suitable time. Plasmid pSV(2)-neo/HPV16 was transfected into cultured kerotinocytes using FuGENE(TM)6. Reverse-transcriptase polymerase chain reaction (RT-PCR) was used to detect c-jun, c-fos and MDM2 mRNA in keratinocytes of normal and transfected keratinocytes. RESULTS: After transfection, the morphology and structure of the keratinocytes were changed. The cells became larger and their nuclei became looser. There were particles in the cytoplasm. The expression of HPV16 mRNA was detected in the keratinocytes after transfecting pSV(2)-neo/HPV16 and the product was 110 bp fragment. The DNA fragment (7.9 kb) of HPV16 was also detected. In the transfected keratinocytes, the exprssions of c-jun, c-fos and MDM2 mRNA were detected and the products were 196 bp, 332 bp and 548 bp fragments, respectively. However, the above-mentioned expressions those were not detected in normal keratinocytes. CONCLUSION: HPV leads to cell proliferation by expressing c-jun, c-fos and MDM2.