Infection of oral keratinocytes with human papilomaviruses(HPV) may involved in the development of oral SCC. E6 and E7 gene of HPV can art together to produce efficient immortalization of primary human epithelial cells, providing further evidence for the role HPV in tumorogenesis. It has been necessary to get optimum medium condition and calcium concentration in order to develop the differentiation and proliferation of cultured IHOK in vitro. IHOK can be used in intermediate cell for oral carcinogenesis.
The purpose of this study was to examine optimum medium requirement and calcium on cultured IHOK, and to apply this culture condition to study oral diseases and tumors in future. Caltured NKOK and IHOK were divided into 5 groups as G1(KBM bullet kit), G2(-BPE), G3(-EGF), G4(-Insulin) and G5(-Hydrocortisone) according to medium conditions to get a growth curve, respectively. According to 0.15, and 1.2 mM Ca concentration, cultured NHOK and IHOK were harvested to take growth curve, and TGase activity assay of cultured NHOK and IHOK, respectively. The obtained results were as follows.
1. Cultured IHOK showed most basal like cell, decreased apoptosis and doubling time(about 25hrs).
2. Absence of BPE, EGF showed inhibitory effect on cultured NHOK and IHOK. Especially absence of hydrocortisone showed more inhibitory effect on cultured NHOK and IHOK.
3. Cultured IHOK showed small foci of stratification in 0.15mM and small areas of stratification, flattened epithelial cells with irregular border in 1.2mM.
4. Cultured NHOK showed more TGase 1 activity in all calcium condition than those of cultured IHOK.
5. Cultured IHOK showed less TGase 1 activity in high calcium condition than that of NHOK.
From the aboving results, it was suggested that although optimum calcium concentration of cultured IHOK was similar to cultured NHOK. Cultured NHOK grew relatively well although they showed increased TGase 1 activity in high calcium compared to low calcium.
|