Effect of morpholino antisense oligonucleotide against lumican mRNA in human embryonic kidney (HEK) 293 cells
Effect of morpholino antisense oligonucleotide against lumican mRNA in human embryonic kidney (HEK) 293 cells
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DOI：
10.1111/j.1440-1827.2004.01593.x
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发表时间：
2004-02-01
影响因子：
2.2
通讯作者：
Naito, Z
中科院分区：
文献类型：
--
作者：
Ishiwata, T;Fujii, T;Naito, Z
Lumican是富含小亮氨酸的蛋白多糖（SLRP）家族的一员，在角膜创面愈合、缺血和再灌注心脏以及几种癌症组织中过度表达。Lumican被认为可以调节胶原原纤维直径和纤维间距。然而，lumican对细胞生长的影响尚未得到充分的研究。在本研究中，我们试图阐明lumican是否有助于人胚胎肾（HEK） 293细胞的生长，使用针对lumican mRNA的morpholino反义寡核苷酸（m-anti oligo）。m -抗寡核苷酸是一种新型寡核苷酸，与硫代型寡核苷酸相比，具有更高的反义活性、更高的水溶性和对靶细胞中核酸酶的抗性。用激光共聚焦显微镜观察HEK 293细胞24 h后，荧光素5-异硫氰酸酯（FITC）偶联寡核苷酸在细胞质和细胞核内的表达。m -抗lumican mRNA寡核苷酸强烈抑制HEK 293细胞中lumican蛋白的合成，HEK细胞的生长速度高于对照组。这些结果可能提示lumican蛋白在体外对HEK 293细胞生长具有抑制作用。
Lumican is a member of the small-leucine-rich proteoglycan (SLRP) family and is overexpressed during wound healing of the cornea, in ischemic and reperfused heart, and in several cancer tissues. Lumican is considered to regulate the collagen fibril diameter and interfibrillar spacing. However, the effect of lumican on cell growth has not been adequately examined. In the present study, we attempted to clarify whether lumican contributes to human embryonic kidney (HEK) 293 cell growth, using the morpholino antisense oligonucleotide (m-anti oligo) against lumican mRNA. M-anti oligo is a novel oligonucleotide and exhibits a higher antisense activity, higher water solubility, and greater resistance to nucleases in target cells than phosphorothioate types of oligonucleotide. After delivery of m-anti oligo against lumican mRNA, the fluorescein 5-isothiocyanate (FITC) conjugated oligonucleotides were observed in the cytoplasm and nucleus of HEK 293 cells at 24 h by confocal laser microscopy. M-anti oligo for lumican mRNA strongly inhibited the synthesis of lumican protein in the HEK 293 cells, and the HEK cell growth rate was higher than those in the control groups. These findings may indicate that lumican protein has an inhibitory effect on HEK 293 cell growth in vitro.