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利用激光显微切割技术对人分泌期子宫内膜基因表达进行比较。

Comparison in gene expression of secretory human endometrium using laser microdissection.

作者信息

Yanaihara Atsushi, Otsuka Yukiko, Iwasaki Shinji, Koide Keiko, Aida Tadateru, Okai Takashi

机构信息

Department of Obstetrics and Gynecology, Showa University School of Medicine, Hatanodai, Shinagawa-ku, Tokyo, Japan.

出版信息

Reprod Biol Endocrinol. 2004 Sep 17;2:66. doi: 10.1186/1477-7827-2-66.

Abstract

BACKGROUND

The endometrium prepares for implantation under the control of steroid hormones. It has been suggested that there are complicated interactions between the epithelium and stroma in the endometrium during menstrual cycle. In this study, we demonstrate a difference in gene expression between the epithelial and stromal areas of the secretory human endometrium using microdissection and macroarray technique.

METHODS

The epithelial and stromal areas were microdissected from the human endometrium during the secretory phase. RNA was extracted and amplified by PCR. Macroarray analysis of nearly 1000 human genes was carried out in this study. Some genes identified by macroarray analysis were verified using real-time PCR.

RESULTS

In this study, changes in expression <2.5-fold in three samples were excluded. A total of 28 genes displayed changes in expression from array data. Fifteen genes were strongly expressed in the epithelial areas, while 13 genes were strongly expressed in the stromal areas. The strongly expressed genes in the epithelial areas with a changes >5-fold were WAP four-disulfide core domain 2 (44.1 fold), matrix metalloproteinase 7 (40.1 fold), homeo box B5 (19.8 fold), msh homeo box homolog (18.8 fold), homeo box B7 (12.7 fold) and protein kinase C, theta (6.4 fold). On the other hand, decorin (55.6 fold), discoidin domain receptor member 2 (17.3 fold), tissue inhibitor of metalloproteinase 1 (9 fold), ribosomal protein S3A (6.3 fold), and tyrosine kinase with immunoglobulin and epidermal growth factor homology domains (5.2 fold) were strongly expressed in the stromal areas. WAP four-disulfide core domain 2 (19.4 fold), matrix metalloproteinase 7 (9.7-fold), decorin (16.3-fold) and tissue inhibitor of metalloproteinase 1 (7.2-fold) were verified by real-time PCR.

CONCLUSIONS

Some of the genes we identified with differential expression are related to the immune system. These results are telling us the new information for understanding the secretory human endometrium.

摘要

背景

子宫内膜在类固醇激素的控制下为着床做准备。有研究表明,月经周期中子宫内膜的上皮细胞和基质之间存在复杂的相互作用。在本研究中,我们使用显微切割和宏阵列技术展示了分泌期人子宫内膜上皮和基质区域基因表达的差异。

方法

在分泌期从人子宫内膜显微切割出上皮和基质区域。提取RNA并通过PCR进行扩增。本研究对近1000个人类基因进行了宏阵列分析。通过宏阵列分析鉴定出的一些基因使用实时PCR进行了验证。

结果

在本研究中,排除了三个样本中表达变化<2.5倍的情况。总共28个基因在阵列数据中显示出表达变化。15个基因在上皮区域强烈表达,而13个基因在基质区域强烈表达。在上皮区域中表达变化>5倍的强烈表达基因有WAP四二硫键核心结构域2(44.1倍)、基质金属蛋白酶7(40.1倍)、同源盒B5(19.8倍)、msh同源盒同源物(18.8倍)、同源盒B7(12.7倍)和蛋白激酶C,θ(6.4倍)。另一方面,核心蛋白聚糖(55.6倍)、盘状结构域受体成员(17.3倍)、金属蛋白酶组织抑制剂1(9倍)、核糖体蛋白S3A(6.3倍)以及具有免疫球蛋白和表皮生长因子同源结构域的酪氨酸激酶(5.2倍)在基质区域强烈表达。通过实时PCR验证了WAP四二硫键核心结构域2(19.4倍)、基质金属蛋白酶7(9.7倍)、核心蛋白聚糖(16.

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