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Expression of CD44 in Cultured Human Trabecular Meshwork Cells

机译：CD44在培养的人小梁网细胞中的表达

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Purpose: To determine whether cultured human trabecular meshwork cells express CD44 and to discuss their possible relationship with primary open angle glaucoma. Methods: Human trabecular meshwork cells were cultured in DMEM/F12 media. Total RNAs from the cells were extracted with Trizol reagent. Messenger RNA expression of CD44 in human trabecular meshwork cells was examined by using reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. Expression of CD44 was confirmed by Western-blotting and immunofluorescent microscopy. Effect of CD44-specific antisense oligonucleotide on adhesion of trabecular meshwork cells to hyaluronate was determined by MTT assay. Results: A single RT-PCR product whose size was 471bp was obtained. A band about 80kD was stained by Western-blot. Immunofluorescent examination of expression of CD44 on the cell surface was positive and reactions were mainly localized in cell membranes. Adhesion of trabecular meshwork cells to hyaluronate was inhibited by CD44-specific antisense oligonucleotide. Conclusions: Cultured human trabecular meshwork cells express CD44. CD44 may play a role in pathogenesis of primary open angle glaucoma.
机译：目的：确定培养的人小梁网细胞是否表达CD44，并讨论它们与原发性开角型青光眼的可能关系。方法：在DMEM / F12培养基中培养人小梁网细胞。用Trizol试剂提取细胞中的总RNA。通过使用逆转录酶-聚合酶链反应（RT-PCR）分析检查人小梁网细胞中CD44的信使RNA表达。通过蛋白质印迹和免疫荧光显微镜检查确认了CD44的表达。通过MTT测定确定CD44特异性反义寡核苷酸对小梁网细胞对透明质酸盐的粘附的作用。结果：获得单个RT-PCR产物，其大小为471bp。通过Western印迹将约80kD的条带染色。免疫荧光检查细胞表面CD44的表达为阳性，反应主要位于细胞膜上。小梁网状细胞对透明质酸的粘附被CD44特异性反义寡核苷酸抑制。结论：培养的人小梁网细胞表达CD44。 CD44可能在原发性开角型青光眼的发病机理中起作用。

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