机构地区:[1]南京医科大学第一附属医院心脏科,210009
出 处:《中华实用儿科临床杂志》2016年第24期1902-1905,共4页Chinese Journal of Applied Clinical Pediatrics
基 金:国家自然科学基金(81370278)
摘 要:目的探索长链非编码RNA(1ncRNA)-uc.167基本生物学特征,及uc.167在小鼠心脏发育过程中的空间分布特征、时间表达规律。方法利用生物信息学软件ENCODE UCSC genomebrowser预测uc.167的基本结构和功能。取不同阶段(孕7.5、11.5、14.5、18.5d)的胚胎小鼠心脏,使用实时荧光定量(RT)-PCR方法检测uc.167及邻近基因在心肌组织中的表达情况,并采用原位杂交技术检测UC.167在心肌组织的分布情况。采用二甲基亚砜方法诱导P19细胞向心肌细胞方向分化,采用RT-PCR方法检测分化过程中(d0、d4、d6、d8、d10)UC.167及邻近基因的表达变化。结果人uc.167全长201bp,定位于基因组5q14.3(chr5:88179623-88179824,GRCh37/hgl9),根据lncRNA分类原则,属Intronic antisense lncRNA;uc.167主要表达于心室肌组织内;uc.167在小鼠胚胎发育过程的表达水平逐渐下调(7.5d:1.000±0.100,11.5d:0.714±0.107,14.5d:0.393±0.043,18.5d:0.125±0.013),而Mef2c的表达水平逐渐上调(7.5d:1.081±0.118,11.5d:2.340±0.351,14.5d:3.958±0.542,18.5d:9.361±0.722),二者呈相反趋势;在P19细胞向心肌细胞分化的过程中,UC.167呈现“先升后降”的表达模式,在胚胎样小体形成(d4)时表达水平达到最高,而后逐渐下调(d0:1.071±0.117,d4:4.714±0.501,d6:3.572±0.414,d8:2.550±0.314,d10:0.786±0.085),临近基因Mefee的表达变化与其呈相反趋势(d0:1.012±0.041,d4:0.353±0.037,d6:2.470±0.329,d8:6.706±0.682,d10:7.765±0.705)。结论UC.167可能在胚胎小鼠心脏发育中具有一定功能;HC.167可能通过负向调控其邻近基因Mef2c来发挥作用。Objective To explore the basic biological characteristics of lncRNA- uc. 167,and its spatial dis- tribution, temporal expression pattern during the mouse embryonic development. Methods The UCSC genome browser of ENCODE was used to analyze preliminary bioinformatics of lneRNAs. Real - time (RT) - PCR was applied to detect the expression of ue. 167 and neighboring genes in the embryonic mouse heart in different stages (PT. 5, P11.5, P14.5, P18.5). Dimethyl sulphoxide was used to induce P19 cell differentiation into the cardiomyocytes. RT -PCR was applied to detect the expression changes in uc. 167 and neighboring genes on differential day 0,4,6,8 and 10. Results Full - length of human he. 167 was 201 bp,and human uc. 167 was located in the genome 5q14.3 ( chr5:88179623 - 88179824,GRCh37/hg19). uc. 167 mainly expressed in the ventrieular muscle tissue. The expression of ue. 167 was gradually decreased in the mouse embryonic heart development process( PT. 5:1. 000 ± 0. 100, P11.5:0. 714± 0. 107, P14.5:0. 393± 0. 043, P18.5:0. 125 ± 0. 013 ) , while the expression of its neighboring Mef2c gene was gradually in- creased( PT. 5:1. 081 ±0. 118 ,Pll. 5:2. 340 ± 0. 351 ,P14.5:3. 958 ±0. 542,P18.5:9. 361 ±0. 722) ,which showed an opposite trend. The expression of ue. 167 during P19 cell differentiation into cardiomyocytes showed a " an increase at first and then a decrease" pattern, and the highest level expression of uc. 167 was on differential day 4 (d0:l. 071 ± 0.117, d4 : 4. 714 ± 0.501, d6 : 3. 572 ± 0.414, d8 : 2. 550 ± 0.314, dl0 : 0. 786 ± 0. 085 ). The expression of neigh- boring gene Mef2c was in an opposite trend( d0:l. 012 ± 0. 041 ,d4: 0. 353 ± 0.037 ,d6: 2. 470 ± 0. 329 ,d8: 6. 706 ± O. 682, d 10: 7. 765 ± O. 705 ). Conclusions It is suggested that uc. 167 may take part in the process of embryonic heart development and may play a role throuah negatively rezulating itsneighboring gene Mef2c.
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