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利用DNA微阵列分析牙周膜和牙龈来源的猪上皮细胞中差异表达基因

Profiling of differentially expressed genes in porcine epithelial cells derived from periodontal ligament and gingiva by DNA microarray.

作者信息

Kurashige Yoshihito, Saitoh Masato, Nishimura Michiko, Noro Daisuke, Kaku Tohru, Igarashi Seiji, Takuma Taishin, Arakawa Toshiya, Inoue Takashi, Abiko Yoshihiro

机构信息

Division of Pediatric Dentistry, School of Dentistry, Health Sciences University of Hokkaido, Hokkaido, Japan.

出版信息

Arch Oral Biol. 2008 May;53(5):437-42. doi: 10.1016/j.archoralbio.2007.12.005. Epub 2008 Feb 1.

Abstract

OBJECTIVE

It is unknown which genes are differentially expressed in cultured epithelial cells derived from the epithelial rests of Malassez (ERM) in periodontal ligament and oral gingival epithelium (OGE). This study analysed the different gene expression of OGE and ERM cells using a DNA microarray technique.

DESIGN

Epithelial cells from ERM and OGE were isolated from porcine periodontal ligament and oral gingival epithelium. Each RNA sample extracted from the cells was reverse transcribed into cDNA and labelled with either cytidine 5-dUTP (Cy5) or cytidine 3-dUTP (Cy3). These labelled cDNA probes were then mixed and simultaneously hybridised to the Pig 13K microarray plate bearing 13,295 different genes (Operon, AL). Cellular enzyme-linked immunosorbent assay (CELISA) was performed to confirm the expression at protein level.

RESULTS

There were nine genes common to the triplicate microarrays in ERM cells and one in OGE cells. Four of the nine genes including tissue factor (TF), FAT cadherin (FAT) and two unknown genes were expressed at levels more than threefold higher in ERM cells than in OGE cells. The protein levels of both TF and FAT in ERM cells were significantly higher than those in OGE.

CONCLUSION

TF and FAT may act as markers to distinguish ERM cells from OGE cells in vitro.

摘要

目的

目前尚不清楚在牙周膜中源自马拉瑟上皮剩余(ERM)的培养上皮细胞和口腔牙龈上皮(OGE)中哪些基因存在差异表达。本研究使用DNA微阵列技术分析了OGE和ERM细胞的不同基因表达。

设计

从猪的牙周膜和口腔牙龈上皮中分离出ERM和OGE的上皮细胞。从细胞中提取的每个RNA样本逆转录成cDNA,并用5-胞苷三磷酸(Cy5)或3-胞苷三磷酸(Cy3)进行标记。然后将这些标记的cDNA探针混合,并同时与带有13295个不同基因(Operon,AL)的猪13K微阵列板杂交。进行细胞酶联免疫吸附测定(CELISA)以确认蛋白质水平的表达。

结果

ERM细胞的三次重复微阵列中有9个基因相同,OGE细胞中有1个基因相同。9个基因中的4个,包括组织因子(TF)、FAT钙黏蛋白(FAT)和2个未知基因,在ERM细胞中的表达水平比在OGE细胞中高3倍以上。ERM细胞中TF和FAT的蛋白质水平均显著高于OGE细胞。

结论

TF和FAT可能作为体外区分ERM细胞和OGE细胞的标志物。

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