Tissue differential microarray analysis of dexamethasone induction reveals potential mechanisms of steroid glaucoma
Tissue differential microarray analysis of dexamethasone induction reveals potential mechanisms of steroid glaucoma
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DOI：
10.1167/iovs.02-0444
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发表时间：
2003-02-01
影响因子：
4.4
通讯作者：
Borrás, T
中科院分区：
文献类型：
--
作者：
Lo, WR;Rowlette, LL;Borrás, T
目的.鉴定人小梁网（HTM）特异性的肌钙蛋白（TIGR/MYOC）特性。目的寻找在地塞米松（DEX）诱导的HTM细胞中高表达而在其他组织中几乎不表达或不表达的基因。通过北方和Western印迹分析，在HTM、人脐静脉内皮细胞、HeLa细胞和人胚胎骨骼肌细胞和汇合时的视神经乳头（ONH）星形胶质细胞中分析DEX（10（-7）M，8-10天）对TIGR/MYOC的诱导。在用过表达野生型和突变形式的TIGR/MYOC的腺病毒感染细胞后，分析加工和分泌。使用Affytron U95 Av 2基因芯片（n = 6）和软件比较HTM、HTM-DEX、ONH星形胶质细胞和ONH星形胶质细胞-DEX的成对表达谱。通过选择具有最高倍数变化值（大于或等于20）的基因来鉴定HTM-DEX特异性基因（与ONH星形胶质细胞-DEX相比）。通过使用基因数据库，将倍数变化值为4或更多的基因与青光眼相关的位点相匹配。DEX诱导的TIGR/MYOC仅发生在HTM细胞中。不同细胞类型的分泌和糖基化特征保持相同。表达谱分析揭示了HTM-DEX中多个差异上调的基因，除了TIGR/MYOC外，还包括丝氨酸蛋白酶抑制剂（α（1）-抗胰凝乳蛋白酶）、神经保护因子（色素上皮衍生因子）、抗血管生成因子（角膜衍生转录物6）和前列腺素合酶（前列腺素D-2合酶）。在249个基因中，有15个基因的倍数变化值为4或更高，定位于与大肠癌连锁的基因座。DEX诱导TIGR/MYOC是HTM特异性的，而其分泌和糖基化特征是普遍存在的。HTM-DEX特异性基因的已知功能揭示了DEX治疗期间IOP调节的保护性和损伤性机制的存在。除了TIGR/MYOC之外，其他HTM-DEX特异性基因可能是与青光眼相关的良好候选基因。
PURPOSE. To identify myocilin (TIGR/MYOC) properties that are specific to the human trabecular meshwork (HTM). To search for genes highly expressed in dexamethasone (DEX)-induced HTM cells that are barely expressed or absent in DEX-induced cells from other tissues.METHODS. TIGR/MYOC induction by DEX (10(-7) M for 8-10 days) was analyzed by Northern and Western blot analyses in HTM, human umbilical vein endothelial cells, HeLa cells, and human embryonic skeletal muscle cells and optic nerve head (ONH) astrocytes at confluence. Processing and secretion were analyzed after the cells were infected with adenoviruses over-expressing wild-type and mutant forms of TIGR/MYOC. Affymetrix U95Av2 GeneChips (n = 6) and software were used to compare paired expression profiles of HTM, HTM-DEX, ONH astrocytes, and ONH astrocytes-DEX. Identification of HTM-DEX-specific genes (compared with ONH astrocytes-DEX) was performed by selecting genes with the highest fold change values ( greater than or equal to20). Genes with fold change values of four or more were matched with loci linked to glaucoma, by using gene databases.RESULTS. TIGR/MYOC induction by DEX occurred only in HTM cells. Secretory and glycosylation characteristics remained the same across cell types. Expression profile analysis revealed multiple genes differentially upregulated in HTM-DEX including, in addition to TIGR/MYOC, a serine protease inhibitor (alpha(1)-antichymotrypsin), a neuroprotective factor (pigment epithelium-derived factor), an antiangiogenesis factor (cornea-derived transcript 6), and a prostaglandin synthase (prostaglandin D-2 synthase). Fifteen of the 249 genes with fold change values of four or more mapped to glaucoma-linked loci.CONCLUSIONS. The induction of TIGR/MYOC by DEX is HTM-specific, whereas its secretory and glycosylation characteristics are ubiquitous. The known functions of HTM-DEX-specific genes reveal the presence of protective and damaging mechanisms for regulation of IOP during DEX treatment. Besides TIGR/MYOC, other HTM-DEX-specific genes may be good candidates for linkage to glaucoma.