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Transcriptional expression in human periodontal ligament cell driven by orthodontic force : RNA-sequencing study

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 교정력에 의한 사람 치주인대 세포의 유전자 발현 : RNA-sequencing 연구 
 College of Dentistry (치과대학) 
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본 연구의 목적은 교정력을 적용했을 때, 치주인대 세포에서의 유전자 발현의 변화를 RNA sequencing을 통해 알아보는 것이다. 교정 진단 결과, 발치가 필요한 10명의 환자를 대상으로 연구를 진행하였다. 치주인대 세포가 물리적 자극을 받는 상황을 시뮬레이션 하기 위해 실험군에 교정력을 가하였고, 대조군은 힘을 가하지 않은 채로 두었다. 발치한 세포로부터 치주인대 세포를 분리한 후, geneset enrichment analysis (GSEA), Differential Expressed Gene (DEG) analysis 및 real time PCR 분석을 실시하였다. GSEA 결과 대부분의 유전자 세트들은 세포 주기와 관련되어 있었고, DEG 분석을 통해 13개의 up-regulated genes 과 20개의 down-regulated gene을 찾았다. Real time PCR을 통해 5개의 up-regulated genes 과 6개의 down-regulated genes 이 RNA sequencing 결과와 일치함을 발견하였다. 유전자 기능 탐색을 통해 최종적으로 치주인대 세포의 재생과 관련된 3개의 유전자(CPNE3, OPHN1 and PPM1F)를 찾았다. 교정력이 가해졌을 때, 치주인대 세포 내 CPNE3 및 OPHN1 유전자는 up-regulated 되며, PPM1F 유전자는 down-regulated 된다. 후속적인 연구(예를 들면, knock-out model을 이용한 functional study)를 통해 이러한 3개의 유전자가 직접적으로 치주인대 세포 재생에 영향을 미치는 것을 확인하는 것이 필요하다.

The aims of this study were to investigate changes in gene expression of periodontal ligament (PDL) cells after mechanical stimulus and to establish mechanism (signaling pathway) through RNA sequencing. We selected ten patients who required tooth extraction for orthodontic treatment. To stimulate the PDL, orthodontic force was applied to the first premolar for three weeks (the experimental group; n=10), while the first premolar on the other side were left untreated (the control group; n=10). After the PDL cells were isolated from the extracted teeth, gene set enrichment analysis (GSEA), differential expressed gene (DEG) analysis, and real time PCR were performed between the experimental and control groups. GSEA demonstrated that most of the gene sets were related to the cell cycle pathway. In the experimental group, 13 up-regulated genes and 20 down-regulated genes were found compared to the control group through DEG analysis. Real time PCR results confirmed that five up-regulation genes and six down-regulated genes were consistent with RNA sequencing results. We finally ruled out three genes (CPNE3, OPHN1, and PPM1F) which is related periodontal ligament regeneration by these analyses. After mechanical stimulus, CPNE3 and OPHN1 genes are up-regulated and PPM1F gene is down-regulated in periodontal ligament cells. We found the three genes can directly affect periodontal ligament, and further studies are needed to clarify the relationship between mechanical stimulus and periodontal ligament cells.
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2. College of Dentistry (치과대학) > Dept. of Dental Education (치의학교육학교실) > 3. Dissertation
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