二羟环氧苯并芘恶性转化细胞的microRNA表达谱  

MicroRNA profiles of malignantly transformed cells induced by anti-benzo-a-pyrene-trans-7,8-dihy- drodiol-9,110-epoxide

作　　者：蒋义国[1] 沈月兰[1] 付娟[1] 周兰兰[1] 曹虹[2] 

机构地区：[1]广州医学院化学致癌研究所与预防医学系,510182 [2]江西省人民医院普通外科

出　　处：《中华劳动卫生职业病杂志》2008年第2期81-85,共5页Chinese Journal of Industrial Hygiene and Occupational Diseases

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（BPDE）恶性转化细胞的microRNA（miRNA）表达谱，寻找恶性转化细胞与对照细胞差异表达的miRNA。方法以溶剂二甲亚砜助溶的终浓度为2．0μmol／L的BPDE培养液培养人支气管上皮细胞株16HBE，获得BPDE恶性转化的细胞模型为实验组，同时，以含二甲亚砜的培养液培养正常16HBE作为对照组。通过微阵列技术检测实验组与对照组327个人类miRNA，并选择差异表达明显的miR-10a和miR-320用反转录荧光定量PCR方法对微阵列结果加以验证。结果获得2组细胞327个miRNA表达谱，发现差异表达miRNA55个，其中46个表达上调，9个表达下调，并得到反转录荧光定量PCR的验证。结论获得BPDE恶性转化细胞与正常16HBE细胞差异表达的miRNA，为进一步寻找BPDE恶性转化细胞特征性miRNA提供了研究基础。Objective To screen microRNA （miRNA） profiles of malignantly transformed cells induced by anti-benzo-a-pyrene-trans-7,8-dihydrodiol-9, 10-epexide （BPDE） and to look for miRNAs which is expressed differently between malignantly transformed cells and normal human bronchial epithelial cells 16HBE. Methods Experimental group was the malignantly transformed 16HBE which was induced by cultured with final concentration 2.0μmol/L of BPDE which was dissolved in dimethyl salphoxide. The control group was 16HBE that was cultured with minimal essential medium including dimethyl sulphoxide. 327 miRNAs were tested be-tween those two groups with miRNA microarray analysis. MiR-lOa that was downexpressed and miR-320 that was overexpressed were selected to be validated by miRNA specific quantitative real-time reverse transcriptase chain reaction （miR qRT-PCR）. Results 327 human miRNAs were tested with miRNA microarray analysis. 55 miRNAs were found expressing differently between those two groups and of which 46 were overexpressed and 9 were downexpressed. Some data were validated by quantitative RT-PCR. Conclusion miRNAs expressed signifcantly between malignantly transformed 16HBE and normal cells and this helps us look for unique miRNAs of malignantly transformed cells induced by BPDE, but there should have more sufficient evidences to prove their functions in malignant cells.

关 键 词：二羟环氧苯并芘 上皮细胞 细胞恶性转化 miRNA 微阵列分析 

分 类 号：R114[医药卫生—卫生毒理学]