To deterrnine the effects of insulin-like growth factor system in the aging process of the rat submandibular gland, expression pattems of 19f-1, 19f-2 and 19f-1 receptor were investigated by immunohistochem엽따. 1n addition, age-related changes of 19f-1 and 19f-2 concentrations in serum were deterrnined by radioimmunoassay 19f-1, 19f-2 and 19f-1 receptor were principally expressed 띠 the duct cells. 19f-1 receptors were strongly expre잃ed in 2-week-old group but were gradually decreased with advancing age. 19f-1 expressions were the strongest in the excretory and striated ducts of 7-month-old and 17-month-old groups. 19f-1 expressions, however, were weakiy expressed in the cells of granular convoluted tubules of 7-month잉ld ， 17-month-old and 27-month-old groups. 19f-2 expressions were found in some cells around duαs of 2-week-old group but were persisted relatively weak. The concentrations of 19f-1 and 1향2 in serum were the highest in 2-week-old group but were decreased and maintained at low level with advancing age. These results show that all components of 19f system were expressed in the rat submandibular giand in an age-related manner. Serum concentrations of all components of 19f system were not dosely related with the intra-밍and비ar expresslon patterns 까lerefore ， 1양 systerns may play important roles in the aging process of the rat submandibular gland in an autocrine/paracrine manner

Insulin-like growth factor 2 (Igf-2) and H19 genes are closely linked imprinted genes which have a pivotal role in embryogenesis and fetal development. Igf-2 and H19 are coexpressed in tissues of mesodermal, endodermal and neuroectodermal origin. Rat Igf-2 has a complex structure with three promoters and a complicate imprinting mechanism having an exception of biallelic expression in the choroid plexus, leptomeninges, and fetal tissues of neuroectodermal origin. To detect the expression of maternal and paternal alleles of Igf-2 and H19 during orofacial development, fetal and neonatal hybrid rats, obtained from Wistar and Fisher interstrain rat crosses were used. We also detected the promoter-specificity of Igf-2 transcripts by primers selected from P1, P2, and P3 of Igf-2 gene. RT-PCR analysis of Igf-2 and H19 showed the monoallelic expression of Igf-2 from the paternal allele and H19 from the maternal allele in E15.5 to E19.5 orofaciall structures including the maxilla, tongue, and salivary gland. P3 promoters were active in all tested samples, whereas transcripts derived from P2 promoter arised with approximately half of the tested cases and showed variable alternation. P1 promoter was not transcribed in all tested samples. These results suggest that Igf-2 and H19 may be involved in orofacial development and exhibit parent-of-origin monoallelic expression. On the other hand, in orofaciall development, P2 and P3 promoters except for P1 promoter are transcribed with variable alternative transcripts

p16INK4A and p15INK4B tumor suppressor genes are frequently altered in various human tumors. Hypermethylation of the promoter region of p16INK4A and p15INK4B seem to be the major mechanism of inactivation. To determine whether the change in p16INK4A and p15INK4B methylation status occur in oral squamous cell carcinomas (OSCCs) and benign oral epithelial hyperplasias, we analyzed 46 OSCCs and 20 benign oral epithelial hyperplasias by methylation-specific PCR. We also analyzed a subset of the samples for p16INK4A and p15INK4B protein expression by immunohistochemistry. The promoter region of p15INK4B was hypermethylated in 13 specimens of the 15 finally analyzed OSCCs and three specimens of the five analyzed benign oral epithelial hyperplasias. By immunohistochemical analysis, we confirmed the loss of p15INK4B expression of all hypermethylated specimens. The promoter region of p16INK4A was amplified by both an unmethylated- and a methylated-specific primers in just one OSCCs. The remaining specimens including 11 OSCCs and four benign oral epithelial hyperplasias were normally methylated. By immunohistochemistry, we analyzed the loss of p16INK4A expression in seven specimens of the 12 OSCCs and two specimens of the four benign oral epithelial hyperplasias. Except for one OSCC, however, all specimens showing loss of expression were normally methylated. These results suggest that loss of p16INK4A and p15INK4B protein expression play an important role in the development of both OSCCs and benign oral epithelial hyperplasias.