人脂肪干细胞脂向分化中长非编码RNA的差异表达.docxVIP

人脂肪干细胞脂向分化中长非编码RNA的差异表达岳文峻，王香梅，张? 芹，黄海华，韦有万，彭? 智广东医学院附属医院，广东省湛江市? 524000Differentially expressed long non-coding RNAs in adipogenic differentiation of human adipose-derived stem cellsYueWen-jun, Wang Xiang-mei, Zhang Qin, Huang Hai-hua, Wei You-wan, PengZhiAffiliated Hospital of Guangdong Medical University, Zhanjiang 524000, Guangdong Province, China摘要? 背景：肥胖导致了包括高血压、冠心病、脂肪肝、高脂血症、2型糖尿病在内的诸多疾病，因此深入理解脂肪细胞分化机制对于肥胖的防治具有深远意义。目前对于脂向分化机制的研究多集中在微小RNA上，对于长非编码RNA在脂向分化过程中的作用尚且知之甚少。目的：获得脂向分化过程中差异倍数明显的长非编码RNA，并进一步筛选在脂向分化过程中可能发挥重要作用的长非编码RNA进行验证。方法：取人腹部皮下脂肪，采用组织块培养法获取脂肪干细胞，传至第3代后进行成脂诱导分化，通过微阵列技术对脂向分化过程中0，5，12 d长非编码RNA及mRNA差异性表达量进行统计，并结合生物信息学报告筛选出呈现明显差异性表达的长非编码RNA，通过qRT-PCR进行验证。结果与结论：脂向分化过程中以差异倍数1.5(P 0.05)为标准，上调长非编码RNA的数量5 d vs. 0 d 748个，12 d vs. 0 d 847个，12 d vs. 5 d 593个；下调长非编码RNA的数量5 d vs.0 d 828个，12 d vs.0 d 1 113个，12 d vs.5 d 750个；结合生物信息学分析结果，在与脂类代谢有关的28个长非编码RNA中根据诱导0，5，12 d差异表达倍数较高且其靶基因可能是已知的与成脂相关的基因中，筛选出3个长非编码RNA：AK304548、BP216319、DA852857。通过PCR验证结果显示AK304548、BP216319及其靶基因表达量呈现先下调后上调的趋势，与微阵列测序结果相符，预示其在脂向分化过程中起到调控作用。中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程全文链接：窗体顶端关键词：干细胞；脂肪干细胞；脂向分化；长链非编码RNA差异性表达；lncRNAs；生物信息学报告；qRT-PCR； ?? Abstract：BACKGROUND: The obesity has led to a plenty of diseases including hypertension, coronary heart disease, fatty liver, hyperlipidemia, and type 2 diabetes. Therefore, understanding the mechanism of adipocyte differentiation is of far-reaching significance to the prevention and treatment of obesity. For the current studies of the mechanism of adipocyte differentiation pay more attention to microRNA, rather than long non-coding RNAs (lncRNAs).OBJECTIVE: To obtain the lncRNAs whose fold change was apparent during adipogenic differentiation, and to further screen the lncRNAs that possibly play a crucial role in adipogenic differentiation for verification. METHODS: Subcutaneous fat was obtained from human abdomen. Adipose-derived stem cells were collected using tissue culture method. The third passage of adipose-deri