CAJ | 학술논문

目的：对长链非编码RNA（lncRNA）uc.417进行系统的生物信息学分析，预测其可能参与的生物学过程，并初步研究其在棕色脂肪细胞中的表达。方法应用UCSC在线数据库获取lncRNA uc.417的序列、位置，并分析其特征；应用Ensemble数据库预测lncRNA uc.417靶基因。分离C57BL/6J小鼠肩胛部棕色脂肪，胶原酶法消化、接种培养并诱导其向成熟的棕色脂肪细胞分化。采用逆转录PCR法检测lncRNA uc.417，同源异型基因B6（HOXB6）及同源异型基因B7（HOXB7）在棕色脂肪细胞分化过程中的表达变化，采用t检验比较三者在分化过程第0~10天的表达差异。结果lncRNA uc.417在各物种之间高度保守，其序列所在染色体区域存在众多顺式作用元件结合位置以及大量表观遗传调控线索，可能的调控靶标为HOXB6与HOXB7。lncRNA uc.417在分化过程D4天的相对表达量显著高于D2（2.07±0.77比0.89±0.18，t=4.103，P<0.01），分化过程D8天的相对表达量显著高于D6（10.68±3.28比3.12±0.88，t=-4.238，P<0.01）。结论 lncRNA uc.417在棕色脂肪细胞成脂分化过程中表达量逐渐增加，生物信息学分析提示可能通过调控下游靶基因，参与棕色脂肪细胞成脂分化过程。
목적：대장련비편마RNA（lncRNA）uc.417진행계통적생물신식학분석，예측기가능삼여적생물학과정，병초보연구기재종색지방세포중적표체。방법응용UCSC재선수거고획취lncRNA uc.417적서렬、위치，병분석기특정；응용Ensemble수거고예측lncRNA uc.417파기인。분리C57BL/6J소서견갑부종색지방，효원매법소화、접충배양병유도기향성숙적종색지방세포분화。채용역전록PCR법검측lncRNA uc.417，동원이형기인B6（HOXB6）급동원이형기인B7（HOXB7）재종색지방세포분화과정중적표체변화，채용t검험비교삼자재분화과정제0~10천적표체차이。결과lncRNA uc.417재각물충지간고도보수，기서렬소재염색체구역존재음다순식작용원건결합위치이급대량표관유전조공선색，가능적조공파표위HOXB6여HOXB7。lncRNA uc.417재분화과정D4천적상대표체량현저고우D2（2.07±0.77비0.89±0.18，t=4.103，P<0.01），분화과정D8천적상대표체량현저고우D6（10.68±3.28비3.12±0.88，t=-4.238，P<0.01）。결론 lncRNA uc.417재종색지방세포성지분화과정중표체량축점증가，생물신식학분석제시가능통과조공하유파기인，삼여종색지방세포성지분화과정。
Objective To predict the biological process in which long non-coding RNA (lncRNA) uc.417 might be involved by a series of bioinformatics analysis, and detect its expression in brown adipocytes, so as to lay foundations and provide theoretical basis for the further studies of lncRNA uc.417 biological functions in brown preadipocytes and obesity. Methods The sequence, position and structure characteristics of lncRNA uc.417 was acquired from UCSC database, and target genes of lncRNA uc.417 were predicted by Ensemble database. The brown preadipocytes were removed, collagenase digestied, incubated and induced to differentiate into mature brown fat cells from C57BL/6J mice. The expression of lncRNA uc.417, homeotic gene B6 (HOXB6) and homeotic gene B7 (HOXB7) were dected using real-time quantitative polymerase chain reaction(PCR). Student′s t test was used to compare their expression level during the differention. Results LncRNA uc.417 had highly conserved property among different species. The chromosomal regions of lncRNA uc.417 had many cis element binding locations and epigenetic regulation clues, and the possible candidate target genes were HOXB6 and HOXB7. The relative expression of lncRNA uc.417 in differentiation day 4 was significantly higher than day 2 (2.07±0.77 vs 0.89±0.18 , t=4.103, P<0.01) , day 8 was significantly higher than day 6 (10.68 ± 3.28 vs 3.12 ± 0.88, t=-4.238, P<0.01). Conclusion The expression of lncRNA uc.417 in brown adipocytes increases during the differentiation. It may play a crucial role in the process of brown preadipocyte-adipocyte transition via regualting the downstream target genes.