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培养的人视网膜色素上皮细胞中表达的转化生长因子-β的鉴定。

Identification of transforming growth factor-beta expressed in cultured human retinal pigment epithelial cells.

作者信息

Tanihara H, Yoshida M, Matsumoto M, Yoshimura N

机构信息

Department of Ophthalmology, Faculty of Medicine, Kyoto University, Japan.

出版信息

Invest Ophthalmol Vis Sci. 1993 Feb;34(2):413-9.

PMID:8440596
Abstract

PURPOSE

This study examined whether retinal pigment epithelial (RPE) cells have the capacity to express transforming growth factor-beta (TGF-beta). Also examined were TGF-beta isoform genes expressed in the RPE cells.

METHODS

Complementary DNA (cDNA) was generated from polyA+ RNA extracted from human RPE cells in culture, and polymerase chain reaction (PCR) using degenerate and specific primers of the known TGF-beta isoforms was carried out by using the cDNAs as templates. Sequencing and Southern blot analysis of the PCR products, and Northern blot analysis were performed to identify which isoforms are expressed in RPE cells.

RESULTS

PCR using degenerate primers showed that most of the amplified sequences were derived from TGF-beta 2, although TGF-beta 1 expression was shown by using specific primers. Northern blot analyses confirmed not only the expression of the TGF-beta gene, but also suggested that alternative splicing of TGF-beta mRNA in the RPE cells occurred.

CONCLUSION

Cultured human RPE cells expressed TGF-beta 1 and beta 2 genes, whereas gene expression of TGF-beta 3 was not confirmed. Our data suggest that RPE cell are intraocular origin of TGF-beta production.

摘要

目的

本研究检测视网膜色素上皮(RPE)细胞是否具有表达转化生长因子-β(TGF-β)的能力。同时检测了RPE细胞中表达的TGF-β亚型基因。

方法

从培养的人RPE细胞中提取的polyA + RNA生成互补DNA(cDNA),并以cDNA为模板,使用已知TGF-β亚型的简并引物和特异性引物进行聚合酶链反应(PCR)。对PCR产物进行测序和Southern印迹分析,并进行Northern印迹分析,以鉴定RPE细胞中表达的亚型。

结果

使用简并引物的PCR表明,大多数扩增序列来自TGF-β2,尽管使用特异性引物显示了TGF-β1的表达。Northern印迹分析不仅证实了TGF-β基因的表达,还表明RPE细胞中TGF-βmRNA发生了可变剪接。

结论

培养的人RPE细胞表达TGF-β1和β2基因,而未证实TGF-β3的基因表达。我们的数据表明RPE细胞是眼内TGF-β产生的来源。

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