Dentin is a mineralized tissue formed by odontoblasts that are differentiated from ectomesenchymal cells , The molecul ar mech anism of odontoblast diffe rentiation remains unclear, Amino acid transporters play an important role in s up plying nutri tion to normal a nd ca ncer cells including odntoblasts, and for cell proliferation , Amino acid transport system L is a maj or nutrient t ransport system responsible for the Na+' -independent transport o[ neutral amino acids incJuding several essentiaJ amino acids , The system L is divided into two major subgroups, the L-type amino acid transporter 1 (LAT1) and the L-type amino acid transporter 2 (LAT2) , In this study, the expression pattern and role of amino acid transport system L were, therefore, investigated in the differentiation of MDPC-23 cells derived from mouse dental papilla celJs , To determi ne the expression Jevel o[ amino acid transport system L participating in intracelJ ular transport of amino acids in the differentiat ion 0 1' MDPC-23 cells, it was examined by RT-PCR, observation of cell morphoJogy‘ A1izaline red-S staining ancl uptake analysis after inclucing experimental differentiation in MDPC-23 cells The res ults were as follows , The LAT1 mRNA was expressed in the early stage of MDPC-23 cell differentiation , The expression leveJ was gradually increased by time course and it was decreased after the late stage, The LAT2 mRNA was not observed in the earJy stage of MDPC-23 cell differentiation, The LAT2 mRNA was expressed at the 11 days 0 1' MDPC-23 cell differentiation and the expression level was gradually decreased by time course, There was no changes in the expression level of 4F2hc mRNA, the cofactor of LAT1 and LAT2, during the differentiation of MDPC-23 cells , The expression of ON mRNA was graduaJJy decreased but the expression of ALP mRNA was increased during differentiation of MDPC-23 cells , The L-Ieucine uptake was increased by time cour se from the early stage to the 9 days in MDPC-23 cell differentiation , The amount of L-Ieucine uptake was maintained to the 11 and 14 days of MDPC-23 cell differentiation As the resul ts‘ it is considered that among neutral amino acid transport system L in differentiation of MDPC-23 cells , the LATl has a key role in cell proliferation in the early stage and middle stage of cell differentiation and the LAT2 has an important roJe in ceJJ differenti ation and mineralization in the Jate stage of cell differentiation for providing cells with neutral a mino acids incJuding several essentiaJ amino acids
The periodontal ligament (PDL) is that soft, specialized connective tissue situated between the cementum covering the root of the tooth and bone forming the socket wall. The PDL is a connective tissue particularly well adapted to its principal function, supporting the teeth in their sockets and at the same time permitting them to withstand the considerable force of mastication. During the life time, PDL is usually exposed to mechanical stress by mastication. However, little is known about the gene which is related to the mechanical stress in PDL. UNC-50 (PDLs22) was identified and isolated from D. melanogater and C. elegance. This gene was also regulated in sensory bristle for mechanotransduction in D. melanogaster. In this study, to uncover the relationship between UNC-50 and mechanical stress, we induced the mechanical stress by medium displacement in cementoblast cell line. After mechanical stress induction UNC-50 expression was analyzed by RT-PCR, Real-time PCR, and western analysis. The expression of UNC-50 was increased after medium displacement of cementoblast in vitro. Collagen type I, type III, and osteonection mRNAs were also strongly expressed after mechanical stress induction. The results of this study suggest that UNC-50 might responsible for molecular event in PDL inducing cementoblast under mechanical stress.
Nuclear factor 1 (NFI) was discovered as a protein required for adenovirus DNA replication in vitro, but it is now clear that NFI protein plays an important role in the expression of many cellular genes. NFI-C null mice demonstrated aberrant odontoblast differentiation, abnormal dentin formation, and thus molar lacking roots while other tissues/or gans in the body, including ameloblasts appear to be unaffected and normal. However, little is known about the mechanism of NFI -C function in odontoblast differentiation and dentin formation. In this study, in order to elucidate the molecular mechanisms of odntoblast differentiation, we examined morphological characteristics of the aberrant odontoblast in NFI-C null mice. we also evaluate the expression of dentin sialophosphoprotein (DSPP) and bone sialoprotein (BSP) mRNAs in the MDPC-23 cells by northern analysis after over-expression and inactiγation of NFI -C into mouse MDPC-23 cells Odontoblasts of the NFI-C null mouse were round in shape, lost their polarity, organized as a sheet of cells, and trapped in osteodentin-like mineralized tissue. Abnormal odontoblasts of NFI-C null mouse revealed the absence of an intercellular junctional complex known as the t erminal webs. MDPC-23 cells started to express DSPP mRNA beginning from the postnatal day of 14 and showed a steady increase as differentiating into odontoblasts. Over-expression of NFI -C increased the expression of DSPP mRNA. Inactivation of NFI - C induced BSP mRNA expression. These results suggest that NFI-C plays an important role in odontoblast differentiation in a cell-type specific manner and thus in dentin formation
Periodontalligament (PDL) fibroblasts have an ectomesenchymal origin and are known to participate not only in formation of PDL but also in the repair and regeneration of the a이acent alveolar bone and cementum. However, little is known about the molecular mechanism which is related to the development and differentiation of PDL cells. Recendy, we reported the PDLs (a periodontalligament-specific) 22 as a PDL fibroblast-specific mRNA which is not expressed in gingival fibroblasts. In this study, to examine the expression and functional characterization of PDμ22 mRNA and prαein in development and differentiation of periodontal 따sue , we carried out northem analysis, insitu hybridization, immunofluorescence and immunohistochemistry. The expression of PDLs22 mRNA was increased with PDL cell differentiation from the confluent to multilayer stage but decreased slighdy with mineralized nodule formation in vitro. πle PDLs22 protein was localized on the nuclear membrane and expressed throughout the differentiation of PDL fibroblasts in vitro. The PDLs22 mRNA and protein were expressed in the differentiating cementoblasts, PDL fibroblasts and osteoblasts along the r∞t surface and alveolar bone of the developing rat teeth. These results indicate that the PDLs22 plays an irnportant role in the differentiation of cementoblasts and osteoblasts and thus homeostasis of cementum, PDL and alveolar bone.
Nitric oxide (NO) plays a key role in the processes of inflammation and carcinogenesis. Three isoforms of NO 야mthase have been identified: endothelial 띠띠c oxide 와nth앓e (NOS), neuronal NOS, and inducible NOS (이OS). The purpose of this study was to investigate the characteristics of iNOS expression in 7, 12-dimethylbenz[alanthracene (DMBA)-induced hamster buccal pouch carcinogenesis. Sixty three outbred young (6-week-old) male Syrian golden hamsters were randomly divided into three groups: DMBA treated group (n=57) and non-treated (n=3), and mineral-oil treated group (n=3). No iNOS activity could be detected in the untreated or mineral oil-treated pouches. 80th cytoplasmic and nuclear stainings were observed in the DMBA-treated pouch kera띠lCX까es. There were iNOS expression 외so in the strorna1 cells. The mean values of iNOS expression in the epithelium increased gradually from control to dysplastic lesions and more to invasive squ따nous cell carcinoma. πle clifference between iNOS expr'않sion in the normal and that the dysplastic and carcinomatous lesions is statistically significant. The mean values of iNOS expression in the stroma increased gradually from control to dysplastic lesions and more to invasive squamous cell carcinoma. The difference between iNOS expression in the normal and that the carcinomatous lesions is statistica11y si맑, ificant. In conclusion, this study has demonstrated that iNOS is expressed in DMBA-induced hamster pouch carcinomas. πlis finding suggests that iNOS expression may be associated with the development of chemically induced oral carcmomas.
Arrùno acid transpoπers play an important role in supplying nutrition to cells and for cell proliferation. System L is a major nutrient transport system responsible for the Na+-independent transport of large neutral amino acids including several essential amino acids. In malignant tumors, a system L transporter L-type amino acid transporter 1 (LATl) is upregulated to support tumor cell growth. In the present study, we have examined the expression and function of system L amino acid transporter in FaDu human pharyngeal squamous carcinoma cells. RT-PCR, real-time quantitative RT-PCR and westem blot analysis have revealed that the FaDu cells express LATl together with its associaω19 protem 4F2hc, whereas the FaDu cells do not express the other system L isoform L-type amino acid transporter 2 (LAT2). 까le uptake of L-(14Clleucine by FaDu cells is Na+-independent and almost completely inhibited by system L selective inhibitor 2-aminobicyclo-(2,2,1)-heptane-2- carboxylic acid (BCH). The profiles of the inhibition of L-[I4Cllellcine uptake by variolls amino acids in the FaDu cells are comparable with those for the LA T1 expressed in Xenopus 。()(찌es. π1e majority of L-[I4Clleucine uptake is, therefore, mediated by LAT1 in the FaDu cells. These results suggest that the transport of neutral amino acids including several essential amino acids in the FaDu human pharyngeal squamous carcinoma cells mediated by LAT1. In addition, specific inhibition of LAT1 by such agents as BCH in pharyngeal squamous cell carcinomas will be a new rationale for anti-cancer therapy.