长链非编码RNA H19及胰岛素样生长因子2在苯并芘诱导C57BL/6J小鼠腭裂发生过程中的动态表达

Mechanism of Long Non-Coding Rna h19 and Insulin-Like Growth Factor 2 Gene in Development of Benzo(a)Pyrene-Induced Cleft Palate in Mice

目的探讨长链非编码RNA H19及胰岛素样生长因子2IGF2在苯并芘BaP诱导C57BL/6J小鼠腭裂发生过程中的动态表达。方法将36只C57BL/6J孕鼠随机分为染毒组和对照组,在胚胎第1d(Embryonic day1,E1)开始染毒,染毒组小鼠以剂量为100 mg/kg的BaP连续灌胃11d,1次/天,在E12时停止灌胃;对照组小鼠以相同剂量的玉米油灌胃。取E13、E14和E15时胎鼠的腭组织,采用实时荧光定量聚合酶链反应技术(RT-PCR)检测不同时期胎鼠腭组织中lncRNA H19及IGF2基因的表达情况。E18时采用苏木精-伊红染色HE对胎鼠腭组织进行形态学观察。统计学方法采用双侧t分布检验及单因素方差分析。结果E13、E14和E15时,染毒组胎鼠腭组织中lncRNA H19基因表达水平总趋势持续上升,其表达量分别是各自对照组的(1.21±0.38)倍,t=0.43,p>0.05;(4.07±1.01)倍,t=6.21,p<0.05;(5.22±0.56)倍,t=8.53,p<0.01。IGF2基因表达水平总趋势持续下降,其表达量分别是各自对照组的(7.45±0.64)倍,t=13.47,p<0.01;(5.39±0.86)倍,t=9.18,p<0.05;(2.74±0.48)倍,t=3.64,p>0.05。E18时,对照组胎鼠的两侧腭接触融合;染毒组胎鼠腭的发育出现生长抑制现象,一侧腭突还没有来得及接触到另一侧腭突而出现腭裂现象。结论lncRNA H19可能通过负调控IGF2基因的表达参与BaP诱导C57BL/6J小鼠腭裂发生和发展。

Objective To investigate the mechanism of long non-coding RNA H19(lncRNA H19)and insulin-like growth factor 2(IGF2)mediated development of cleft palate in C57BL/6J mice exposed to benzoapyrene(BaP).Method Thirty-six pregnant C57BL/6J female mice were randomly divided into two groups(18 in each group):control group(interfered with corn oil)and BaP-treated group(treated with 100 mg/kg of BaP).These pregnant mice were administered by gavage from embryonic day 1(E1)to E11,and some of them were sacrificed on E13,E14 and E15 to collected palatal tissues from fetal mice.Real-time quantitative polymerase chain reaction(RT-PCR)was performed to measure expression of lncRNA H19 and IGF2 genes in palatal tissues that collected at different developmental stages.Hematoxylin-eosin staining(HE)was used to observe changes of palatal tissues in morphology on E18.Two-sided t-distribution test was used for statistical analysis.Result Expression of lncRNA H19 gene on E13,E14 and E15 continuously increased in BaP-treated group and the levels of expressed lncRNA H19 gene were higher than those of control group on the corresponding time points[(1.21±0.38)times,t=0.43,p>0.05;(4.07±1.01)times,t=6.21,p<0.05;(5.22±0.56)times,t=8.53,p<0.01].However,expression of IGF2 gene from E13 to E15 continuously declined in BaP-treated group and the levels were respectively(7.45±0.64)(t=13.47,p<0.01),(5.39±0.86)(t=9.18,p<0.05)and(2.74±0.48)(t=3.64,p>0.05)times of those of control group on the corresponding embryonic days.Conclusion LncRNA H19 might be involved in the occurrence and development of BaP-induced cleft palate in C57BL/6J mice by negatively regulating the expression of IGF2 gene.