二羟环氧苯并芘恶性转化细胞miR-10a及其靶基因HOXA1的表达改变  

Expressions of miR-10a and its target gene HOXA1 in BPDE transformed cells

作　　者：沈月兰[1] 蒋义国[1] 周兰兰[1] 付娟[1] 

机构地区：[1]广州医学院化学致癌研究所与预防医学系,广东广州510182

出　　处：《毒理学杂志》2008年第2期81-84,共4页Journal of Toxicology

基　　金：国家自然科学基金资助项目(30571546;30771780);教育部留学回国人员科研启动基金资助项目(2007-24);广东省自然科学基金资助项目(07117550);广东省高校自然科学重点项目(06Z021)

摘　　要：目的检测二羟环氧苯并芘恶性转化细胞miR-10a表达水平,探讨其与靶基因HOXA1癌基因的关系。方法以正常支气管上皮细胞为对照组,二羟环氧苯并芘恶性转化细胞为实验组,用茎环结构逆转录引物逆转录,定量PCR检测两组细胞miR-10a表达水平。运用半定量逆转录-聚合酶链反应(RT-PCR)、定量PCR和流式细胞术间接法检测2组细胞HOXA1 mRNA和蛋白表达水平。结果二羟环氧苯并芘恶性转化细胞miR-10a表达水平是对照组的0.01倍,RT-PCR HOXA1 mRNA是对照组的(3.12±0.23)倍,定量PCR HOXA1 mRNA是对照组的8.75倍,HOXA1蛋白表达是对照组的3.48倍。结论二羟环氧苯并芘恶性转化细胞miR-10a可能反向调控靶基因HOXA1的表达。Objective To investigate the expression of miR-10a in malignantly transformed cells induced by anti-benzo（a） pyrene-trans-7,8-dihydrodiol-9, 10-epoxide （BPDE） and explore the relationship between miR-10a and its target oncogene HOXA1. Methods To quantify miR-10a expression between malignantly transformed cells and normal human bronchial epithelial cells, we employed a highly sensitive technique that uses stem-loop primers for reverse transcription followed by real-time PCR. On the other hand, we examined the expression of HOXA1 by using half quantitative RT-PCR, real-time PCR and flow cytometry. Results In comparison to the corresponding control cells, miR-10a expression was 0.01 times in transformed cells, HOXA1 mRNA expression was 3.12 ± 0.23 times by RT-PCR and 8.75 times by real-time PCR, HOXA1 protein was 3.48 times by flow cytometry. Conclusion MiR-10a might negatively regulate its target gene HOXA1.

关 键 词：二羟环氧苯并芘 恶性转化细胞 miR-10a HOXA1 

分 类 号：R994.6[医药卫生—毒理学]