Chen Wei, Fu Xiao-Bing, Ge Shi-Li, Sun Tong-Zhu, Sheng Zhi-Yong
Key Research Laboratory of Wound Repair, Burn Institute, 304th Clinical Department, General Hospital of PLA, Beijing 100037, China.
Chin J Traumatol. 2006 Apr;9(2):94-9.
To study the differences of gene expression between earlier gestational skin and later gestational skin of rats with the aids of single primer amplification (SPA) and high-density oligonucleotide DNA array to understand the molecular mechanism of scarless healing.
Total RNAs were isolated from fetal rat skin of the scarless (E15) and scar-forming (E18) periods of gestation (term = 21.5 days). The RNAs from earlier gestational skin (EGS) and later gestational skin (LGS) were both reversely transcribed to cDNAs, then labeled with the incorporation of fluorescent dCTP for preparing the hybridization probes by SPA method. The mixed probes were then hybridized to the oligonucleotide DNA arrays which contained 5,705 probes representing 5,705 rat genes. After highly stringent washing, these DNA arrays were scanned for fluorescent signals to display the differentially expressed genes between the 2 groups of skin.
Among 5,705 rat genes, there were 53 genes (0.93 percent) with differentially expressed levels between EGS and LGS groups, 27 genes, including fibroblast growth factor 2 (FGF2) and follistatin were up-regulated (0.47%) and 26 genes were down-regulated (0.46%) in fetal skin during scarless period versus scar-forming period. Higher expressions of FGF2 and follistatin in EGS than those in LGS were also revealed by RT-PCR method.
High-density oligonucleotide DNA array provided a powerful tool for investigating differential gene expression in earlier and later gestational fetal skins. This technology validates that the mechanism of fetal scarless healing is very complicate and the change of many gene expressions is associated with fetal scarless healing.
借助单引物扩增（SPA）和高密度寡核苷酸DNA芯片研究大鼠妊娠早期皮肤与妊娠晚期皮肤基因表达的差异，以了解无瘢痕愈合的分子机制。
从妊娠无瘢痕期（E15）和瘢痕形成期（E18）（足月为21.5天）的胎鼠皮肤中分离总RNA。将妊娠早期皮肤（EGS）和妊娠晚期皮肤（LGS）的RNA都反转录成cDNA，然后通过掺入荧光dCTP进行标记，用SPA法制备杂交探针。然后将混合探针与包含5705个代表5705个大鼠基因的探针的寡核苷酸DNA芯片杂交。经过高度严格洗涤后，扫描这些DNA芯片以获取荧光信号，以显示两组皮肤之间差异表达的基因。
在5705个大鼠基因中，EGS组和LGS组之间有53个基因（0.93%）表达水平存在差异，在无瘢痕期与瘢痕形成期的胎儿皮肤中，包括成纤维细胞生长因子2（FGF2）和卵泡抑素在内的27个基因上调（0.47%），26个基因下调（0.46%）。RT-PCR法也显示EGS中FGF2和卵泡抑素的表达高于LGS。
高密度寡核苷酸DNA芯片为研究妊娠早期和晚期胎儿皮肤中差异基因表达提供了有力工具。该技术证实胎儿无瘢痕愈合的机制非常复杂，许多基因表达的变化与胎儿无瘢痕愈合有关。
