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Published on: July 10, 2014
Fam83h affected adhesion ability and extracellular matrix collagen formation in ameloblasts
Mingmei Meng1, Yongting Xie1,2,3, Liyan Shi1
1State Key Laboratory of Oral Diseases, National Center for Stomatology, National Clinical Research Center for Oral Diseases, Department of Pediatric Dentistry, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China.
Purpose:
Amelogenesis imperfecta (AI) is a group of inherited diseases characterized by enamel defects. Mutation in multiple genes may cause AI. The objective of this study was to investigate the specific roles of Family with sequence similarity 83 member H (FAM83H) in AI and determine its effect on ameloblasts.
Methods:
To investigate the impact of Fam83h on ameloblast gene expression, we utilized ameloblast line HAT-7 cells with knockdown of Fam83h gene for transcriptome sequencing. The experimental cell samples were undergone library preparation and RNA sequencing expreiments. Cell Adhesion test kit was employed to evaluate the influence of Fam83h on HAT-7 cell adhesion ability. RT-qPCR and western blot were conducted to analyze the expression levels of integrin and collagen proteins upon downregulation of Fam83h.
Results:
RNA-seq analysis revealed differential expression of 572 genes in HAT-7 cells with knockdown of Fam83h gene, including 285 up-regulated genes and 287 down-regulated genes. GO and KEGG enrichment analysis indicated that Fam83h could modulate collagen synthesis in ameloblasts as well as affect the focal adhesion pathway. In the cell adhesion assay, we observed a decrease in adhesion ability due to downregulation of Fam83h in HAT-7 cell line. Furthermore, it was found that Fam83h can upregulated extracellular matrix collagen formation in ameloblasts.
Conclusions:
Fam83h may affect the adhesion ability and extracellular matrix collagen formation in ameloblasts, leading to AI.
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