摘要目的 探讨长链非编码 RNA ( long non-coding RNA,lncRNA)致妊娠糖尿病( gestational diabetes mellitu,GDM)巨大儿发生的作用机制.方法 采用实时荧光定量 PCR(qantitative real-timePCR, qRT-PCR)检测CTD-2012K14.6在GDM和正常胎盘组织中的表达量,并进行胎儿体重相关性分析;运用生物信息分析技术确定CTD-2K14.6调控的下游靶基因,并利用过表达及干扰RNA技术改变CTD-2K4.6在人滋养层细胞系中的表达,及其对下游靶基因表达的调控作用.结果 与正常妊娠胎盘组织相比, CTD-2012K14.6在GDM孕妇的胎盘组织中的相对表达量明显上调(1.70 ± 0.63对1.00 ± 0.56,P＜0.01),且与胎儿体重呈正相关,r=0.850,P＜0.01;在线分析CTD-2012K14.6位于chr16:67,549,214-67,563,958,染色体位置分析发现,CTD-2012K14.6位于CTCF基因的内含子内;过表达CTD-2012K14.6可明显降低CTCF核酸与蛋白表达量,而使胰岛素样生长因子Ⅱ(IGF-Ⅱ)核酸与蛋白表达量显著升高;敲除CTD-2012K14.6显著升高CTCF核酸与蛋白表达量,而明显降低IGF-Ⅱ核酸与蛋白表达量.结论 胎盘组织中异常表达的CTD-2012K14.6可能通过调控CTCF,IGF-Ⅱ影响巨大儿发生发展.

abstractsObjective To explore the role of long noncoding RNA ( lncRNA) CTD-2012K14. 6 in the development of gestational diabetes mellitus (GDM) related macrosomia. Methods The quantitative real-time PCR ( qRT-PCR) was performed to measure the expression of CTD-2012K14.6 in placentas of women with or without GDM, and the quantity of CTD-2012K14. 6 expression and its association with fetal weights were analyzed; Bioinformatic analysis was performed to predict the downstream molecules. CTD-2012K14. 6 over-expressing lentiviral and siRNA was constructed in human trophoblastic cell line HTR-8/SVneo cells, qRT-PCR and Western blot (WB) were used to invest its effect in modulating the expression of downstream molecules. Results The expression of CTD-2012K14.6 in GDM placentas was significantly higher than that in normal controls (1.70 ± 0.63 vs 1.00 ± 0.56,t=3.68,P＜0.01), and positively correlated with fetal weight (r=0.8501, P＜0.01); on-line analysis showed that CTD-2012K14.6 was located at chr16:67,549,214-67,563,958, which was located in the intron of CCCTC-binding factor( CTCF); Up-regulating CTD-2012K14.6 could significantly reduce the expression of CTCF mRNA and protein, and increase the expression of insulin-like growth factor-Ⅱ( IGF-Ⅱ) mRNA and protein, while down-regulating CTD-2012K14.6 could significantly increase the expression of CTCF mRNA and protein, and reduce the expression of IGF-ⅡmRNA and protein. Conclusion The CTD-2012K14. 6 may play an important role in the pathogenesis of GDM related macrosomia by upregulating the expression of CTCF and IGF-Ⅱ.