人肝癌细胞辅酶Ⅱ-依赖性视黄醇脱氢/还原酶选择性剪接新亚型的鉴定和分析

Identification and Analysis of a Novel Alternatively Spliced Variant of NADP(H)-dependent Retinol Dehydrogenase/Reductase in HepG2 Cell

目的:鉴定人肝癌细胞HepG2中辅酶Ⅱ-依赖性视黄醇脱氢/还原酶(NRDR)的选择性剪接新亚型DHRS4L1A1,并检测其在不同细胞中的表达情况。方法:采用快速cDNA末端扩增方法,从HepG2中克隆选择性剪接亚型的cDNA序列;通过查找开放阅读框架,预测并分析剪接新亚型的蛋白质序列和功能;用半定量RT-PCR法检测8种不同细胞株中新剪接亚型的相对表达量。结果:DHRS4L1A1全长1 117 bp,为NRDR选择性剪接新亚型;该亚型在卵巢癌细胞SKOV3中的表达量显著高于其他细胞。结论:DHRS4L1A1表达的蛋白属于SDR家族,该亚型可能参与生殖系统相关酶类的代谢。

Objective： To identitify NADP（H）-dependent retinol dehydrogenase/reduetase（NRDR） alternatively spliced variants, and to detect the expression level of this ahematively splicing variant in different cell lines. Methods： Rapid amplification of eDNA ends（RACE）, RT-PCR was used to clone the full length in HepG2 cell line and detect the expression in different eel1 lines. The amino acid sequence of the translated protein of this splicing variant was predicted and analyzed by searching open reading frame. Results： The full-length of DHRS4L1A1 was 1 117 bp. DHRS4L1A1 protein conserved the eoenzyme binding motifs of SDR family at N terminus, substrate binding motifs at C terminus, and pemxisome-targeting signal. Its expression in ovarian cancer cell line SKOV3 was significantly higher than other cells. Conclusion： The translated protein of DHRS4L1A1 belongs to the SDR family, it had close relationship with metabolic pathways of reproductive system.