The Role of HPV16 E6 Protein on Matrix Metalloproteinase 2 and 9 Expressions in Human Embryo Skin Fibroblast Cell / 대한산부인과학회지

ABSTRACT

OBJECTIVE: The purpose of this study was to evaluate the role of HPV16 E6 protein on the expressions of MMP 2 and 9, major proteins associated with tumor invasion and metastasis, in human embryo skin fibroblast cell. METHODS: HPV16 E6 ORF was obtained by PCR from CaSki cell genomic DNA and was cloned into mammalian expression vector, pTARGET. Negative control vector was constructed with self ligated pTARGET. After confirmation of plasmid with color selection and restriction enzyme digestion, pTARGET-E6 and negative control vector were transfected into Detroit 551. On post-transfection third day, supernatant of both transfected cell were harvested and quantitative analysis was carried out with bradford method. Zymographic analyses were run in parallel. Areas of gelatinolytic activity were detected as clear bands on blue background and analyzed with computerized program. RESULTS: Zymogram followed by densitometric analysis was revealed the increased expression of MMP 2 zymogen (72 kDa) and active form (67 kDa) by 10.5% and 36.3%, respectively. On the contrary, MMP 9 expression was decreased by 0.21% on densitometric analysis. CONCLUSION: HPV16 E6 protein has influenced on MMP 2 over-expression.