Identification of RPE65 in transformed kidney cells
Identification of RPE65 in transformed kidney cells
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DOI：
10.1016/s0014-5793(99)00606-7
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发表时间：
1999-06-11
期刊：
影响因子：
3.5
通讯作者：
Crouch, RK
中科院分区：
文献类型：
--
作者：
Ma, JX;Zhang, DC;Crouch, RK
蛋白RPE 65在眼内类维生素A的加工和/或转运中具有重要作用。类维生素A参与细胞分化、胚胎发生和癌发生。由于肾脏是类维生素A代谢的重要部位，因此已经检测了RPE 65在正常肾脏和转化肾脏细胞中的表达。通过逆转录PCR在转化肾脏细胞系包括人胚肾细胞系HEK 293和非洲绿色猴肾细胞系COS-1和COS-7中检测到RPE 65 mRNA。在相同的PCR条件下，在人原代肾细胞或猴肾组织中未检测到。使用RPE 65的单克隆抗体通过Western印迹分析也在COS-7和HEK 293细胞中鉴定了RPE 65蛋白，但在原代肾细胞或肾组织中没有。从HEK 293和COS-7细胞中扩增出含有全长编码区的RPE 65 cDNA。DNA测序结果表明，HEK 293细胞的RPE 65 cDNA与人视网膜色素上皮细胞的RPE 65 cDNA相同。来自COS-7细胞的RPE 65与人RPE 65在核苷酸和氨基酸水平上分别具有98%和99%的序列同一性。此外，RPE 65 mRNA的检测在三个四分之一的肾肿瘤文化分析，包括先天性中胚层肾瘤和肾透明细胞肉瘤。这些结果表明，转化的肾细胞表达这种类维生素A加工蛋白，表明这些转化的细胞可能具有正常肾细胞中不存在的替代类维生素A代谢。(C)1999年欧洲生物化学学会联合会。
The protein RPE65 has an important role in retinoid processing and/or retinoid transport in the eye, Retinoids are involved in cell differentiation, embryogenesis and carcinogenesis. Since the kidney is known as an important site for retinoid metabolism, the expression of RPE65 in normal kidney and transformed kidney cells has been examined, The RPE65 mRNA was detected in transformed kidney cell lines including the human embryonic kidney cell line HEK293 and the African green monkey kidney cell lines COS-1 and COS-7 by reverse transcription PCR, In contrast, it was not detected in human primary kidney cells or monkey kidney tissues under the same PCR conditions. The RPE65 protein was also identified in COS-7 and HEK293 cells by Western blot analysis using a monoclonal antibody to RPE65, but not in the primary kidney cells or kidney tissues. The RPE65 cDNA containing the full-length encoding region was amplified from HEK293 and COS-7 cells. DNA sequencing showed that the RPE65 cDNA from HEK293 cells is identical to the RPE65 cDNA from the human retinal pigment epithelium. The RPE65 from COS-7 cells shares 98 and 99% sequence identity with human RPE65 at the nucleotide and amino acid levels, respectively. Moreover, the RPE65 mRNA was detected in three out of four renal tumor cultures analyzed including congenital mesoblastic nephroma and clear cell sarcoma of the kidney. These results demonstrated that transformed kidney cells express this retinoid processing protein, suggesting that these transformed tells may have an alternative retinoid metabolism not present in normal kidney cells. (C) 1999 Federation of European Biochemical Societies.