Isolation and characterization of cultured human periodontal ligament fibroblast-specific cDNAs
Isolation and characterization of cultured human periodontal ligament fibroblast-specific cDNAs
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DOI：
10.1006/bbrc.2001.4694
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发表时间：
2001-04-20
影响因子：
3.1
通讯作者：
Han, KY
中科院分区：
文献类型：
--
作者：
Park, JC;Kim, YB;Han, KY
控制牙周膜（PDL）成纤维细胞功能的分子机制仍不清楚。我们推测，分化 PBL 成纤维细胞的特征是由牙周组织中邻近牙龈成纤维细胞中未发现的特定基因的改变的扩展来定义的。为了扩展该组，在培养的人 PDL 和牙龈成纤维细胞之间应用消减杂交，以鉴定 PDL 中差异表达的基因。因此，通过同源性搜索、Northern 分析和原位杂交鉴定并表征了五个候选克隆：PDL（牙周膜特异性）5、-17、-22、-25 和-31。尽管这些克隆的 mRNA 由骨髓细胞表达，很少由牙龈成纤维细胞表达，但在 PDL 细胞中检测到最高表达，均匀分布在整个 PDL 中。在五个候选克隆中，我重点关注PDLs17，因为它是一种假设的蛋白质，其生物学功能尚未被报道在数据库中。制备了针对PDLs17肽的多克隆抗血清，并对骨髓中的PDL成纤维细胞、成骨细胞和基质细胞进行染色，但对牙龈成纤维细胞不进行染色。结果表明，PDLs5、-17、-22、-25和-31克隆可作为PDL成纤维细胞特异性标记，并且PDLs17可作为PDL成纤维细胞分化过程中的重要因素。 (C) 2001 年学术出版社。
The molecular mechanisms that control the function of periodontal ligament (PDL) fibroblasts remain unclear. We speculated that the character of differentiating PBL fibroblasts is defined by the altered expansion of specific genes not found in neighboring gingival fibroblasts in the periodontium. To expand this set, subtractive hybridization was applied between cultured human PDL and gingival fibroblasts to identify genes differentially ex-pressed in PDL. Consequently five candidate clones, PDLs (periodontal ligament specific)5, -17, -22, -25, and -31 were identified and characterized by homology search, Northern analysis, and irt situ hybridization. Although the mRNAs of these clones were expressed by bone marrow cells and rarely by gingival fibroblasts, the highest expression was detected in the PDL cells, which were uniformly distributed throughout the whole PDL. Amongst the five candidate clones, me focused on PDLs17, because it is a hypothetical protein whose biological function has not been reported Set in the database. Polyclonal antiserum raised against PDLs17 peptide was made, and stained the PDL fibroblasts, osteoblast-liIie cells and stromal cells in the bone marrow, but not gingival fibroblasts. The results suggest that clones, PDLs5, -17, -22, -25, and -31 may be used as PDL fibroblast-speeific markers, and that PDLs17 could act as an important factor in the differentiation process of PDL fibroblasts. (C) 2001 Academic Press.