摘要
目的探讨辅酶Ⅱ依赖性视黄醇脱氢/还原酶(NADP-dependent retinol dehydrogenase/reductase,NRDR)与神经母细胞瘤发生的关系。方法采用RACE方法从神经母细胞瘤细胞SK-N-SH和SY-5-Y中克隆NRDR全长cDNA,对获得的序列进行生物信息学分析,Northern杂交确定NRDR组织分布,免疫荧光染色鉴定NRDR细胞内定位,杆状病毒蛋白表达系统表达克隆的NRDR,反相高压液相色谱测定NRDR催化活性。结果从神经母细胞瘤细胞中克隆了一种新的NRDR全长cDNA片段(NCB I登录号为DQ344810),序列分析表明NRDR属短链醇脱氢酶家族。结论NRDR定位于细胞过氧化物酶体,具有辅酶Ⅱ依赖性视黄醇脱氢/还原酶活性。神经母细胞瘤细胞中NRDR表达调控的改变,可能与神经母细胞瘤的发生有关。
Objective To identify the NADPH-dependent retinol dehydrogenase/reductase (NRDR) in neuroblastoma cell line and investigate the relationship between NRDR and neuroblastoma carcinogenesis. Methods NRDR cDNA in neuroblastoma cell line SY-5-Y and SK-N-SH was cloned by RACE. The obtained sequence was bioinformatically analyzed. The expression profile of NRDR in tissues was detected by Northern blotting. The cloned NRDR was expressed by baculovirus protein expression system and its enzymatic activity was identified by reverse phase HPLC. The intracellular location of NRDR was determined by immunofluorescence staining. Results A novel NRDR cDNA sequence was cloned from neuroblastoma cell line whose NCBI accession is DQ344810. Sequence analysis indicated that NRDR belonged to short chain alcohol dehydrogenase/ reductase family. Conclusion NRDR is a novel regulator of retinoid metabolism in neuroblastoma. NRDR may be associated with the carcinogenesis of ncuroblastoma.
出处
《医学分子生物学杂志》
CAS
CSCD
2006年第2期90-95,共6页
Journal of Medical Molecular Biology
基金
国家自然科学基金(No.30470396)~~