ABSTRACT
BACKGROUND: beta-catenin plays a pivotal role in hair follicle development and hair growth cycle. OBJECTIVE: The aim of this study was to identify beta-catenin-regulated genes in cultured human hair outer root sheath (ORS) cells. METHODS: Primary cultured ORS cells were transduced with recombinant adenovirus expressing N-terminal truncated beta-catenin (constitutive active form), and beta-catenin-regulated genes were identified. RESULTS: Overexpression of the constitutively active form of beta-catenin led to induction of Sox9 expression at both mRNA and protein levels. To investigate the potential role of Sox9, we made the recombinant adenovirus expressing green fluorescent protein-tagged Sox9, and then transduced into cultured ORS cells. Interestingly, Sox9 induced the expression of keratin 15, increased the proliferation of ORS cells in vitro, and enhanced colony-forming activity. CONCLUSION: Our results suggest that Sox9 is a beta-catenin-regulated gene in ORS cells, and has potential importance in the regulation of hair follicle homeostasis.
Subject(s)
Humans , Adenoviridae , beta Catenin , Electrolytes , Hair , Hair Follicle , Homeostasis , Keratin-15 , RNA, MessengerABSTRACT
PURPOSE: Usefulness of mouse liver S9 fraction was evaluated for the measurement of the metabolites in the in vitro metabolism study of 18F-labeled radiotracers. MATERIALS AND METHODS: Mouse liver S9 fraction was isolated at an early step in the course of microsome preparation. The in vitro metabolism studies were carried out by incubating a mixture containing the radiotracer, S9 fraction and NADPH at 37 degrees C, and an aliquot of the mixture was analyzed at the indicated time points by radio-TLC. Metabolic defluorination was further confirmed by the incubation with calcium phosphate, a bone mimic. RESULTS: The radiotracer [18F]1 underwent metabolic defluorination within 15 min, which was consistent with the results of the in vivo method and the in vitro method using microsome. Radiotracer [18F]2 was metabolized to three metabolites including 4-[18F]fluorobenzoic acid within 60 min. It is likely that the one of these metabolites at the origin of radio-TLC was identical with the one that obtained from the in vivo and in vitro (microsome) method. Compared with the in vitro method using microsome, the method using S9 fraction gave a similar pattern of the metabolites but with a different ratio, which can be explained by the presence of cytosol in the S9 fraction. CONCLUSION: These results suggest that the findings of the in vitro metabolism studies using S9 fraction can reflect the in vivo metabolism of novel radiotracers in the liver. Moreover, this method can be used as a tool to determine metabolic defluorination along with calcium phosphate absorption method.