鸡前脂肪细胞分化过程中BMP2、PDK1和LPL基因3'-UTR的动态变化研究OA
Dynamic Changes in the 3'-UTR Regions of BMP2,PDK1,and LPL Genes during Chicken Preadipocyte Differentiation
旨在鉴定鸡前脂肪细胞(ICP1)分化过程中脂肪生成基因的3'-UTR动态变化.本研究以鸡永生化前脂肪细胞系(ICP1)为试验材料,试验设置0、24、48、72、96和120 h共6个时间点分组.采用160 μmol・L⁻¹油酸诱导细胞分化,通过油红 O 染色观察分化 0和 120 h脂滴沉积并定量分析脂肪生成;利用 qRT-PCR检测成脂标志基因(PPARγ、FABP4)与目的基因(BMP2、PDK1、LPL、LDAH)的 mRNA表达水平;采用 3'-RACE技术及测序验证BMP2和PDK1基因3'-UTR长度变异.所有试验均设置3次生物学重复及相应技术重复.结果表明:在不同分化时期各基因的表达水平均存在差异,其中BMP2和PDK1在分化前期(24～48 h)的基因表达水平明显高于后期,而LPL和LDAH基因表达水平则随细胞分化程度逐渐升高,在48 h时达到最大,随后随着分化时间的增加而逐渐下降;BMP2、PDK1和LPL基因在ICP1细胞分化过程中分别转录出5条、4条和4条不同3'-UTR长度的转录本;此外,通过定量分析发现BMP2和PDK1基因在细胞分化前期(0、24 h)主要表达3'-UTR长转录本,随着分化进行,BMP2在96和120 h时长、短转录本同时存在且表达水平相同,而PDK1在48、72、96和120 h时长、短转录本表达水平相同.进一步分析发现,与0 h相比,48 h时BMP2和PDK1基因倾向于表达3'-UTR更长的转录本.鸡前脂肪细胞分化过程中脂肪生成基因有不同3'-UTR长度转录本的表达,且在分化过程不同3'-UTR的转录本表达具有显著差异.研究结果为深入探讨APA调控基因转录进而影响细胞分化的分子机制提供了新素材和方向.
This study aimed to identify dynamic changes in the 3'-UTRs of adipogenesis genes during the differentiation of chicken preadipocytes(ICP1).In this study,the chicken immortalized preadipocyte cell line(ICP1)was used as the experimental material.Cells were divided into 6 time-point groups(0,24,48,72,96,and 120 h).Adipocyte differentiation was induced with 160 μmol·L-1 oleic acid.Lipid droplet deposition at 0 and 120 h was observed by Oil Red O staining and quantified to evaluate adipogenesis.The mRNA expression levels of adipogenic marker genes(PPARγ and FABP4)and target genes(BMP2,PDK1,LPL,and LDAH)were detected by q RT-PCR.In addition,3'-RACE and sequencing were performed to verify the 3'-UTR length variants of the BMP2 and PDK1 genes.All experiments were conducted with 3 biological replicates and corresponding technical replicates.The results showed that the expression levels of these genes varied at different stages of differentiation.Among them,the expression levels of BMP2 and PDK1 were significantly higher in the early stage of differentiation(24-48 h)than in the later stage,whereas the expression levels of LPL and LDAH gradually increased with cell differentiation,reached a peak at 48 h,and then gradually decreased with prolonged differentiation time.During the differentiation of ICP1 cells,BMP2,PDK1,and LPL generated 5,4,and 4 transcripts with different 3'-UTR lengths,respectively.In addition,quantitative analysis showed that BMP2 and PDK1 mainly expressed long 3'-UTR transcripts at the early stage of differentiation(0 and 24 h).As differentiation progressed,both long and short transcripts of BMP2 were present and expressed at similar levels at 96 and 120 h,whereas PDK1 exhibited similar expression levels of long and short transcripts at 48,72,96,and 120 h.Further analysis indicated that,compared with 0 h,BMP2 and PDK1 tended to express transcripts with longer 3'-UTRs at 48 h.During differentiation,adipogenesis genes with different 3'-UTR long and short transcripts were expressed in chicken preadipocytes,and there was a significant difference in the expression of different 3'-UTRs.The findings provide new insights and directions for exploring the molecular mechanisms by which APA regulates gene transcription and subsequently influences cell differentiation.
黄琴;赵素娟;杜志强
长江大学动物科学技术学院,荆州 434020东北农业大学动物科学技术学院,哈尔滨 150000长江大学动物科学技术学院,荆州 434020
农业科技
鸡前脂肪细胞3'-UTR可变多聚腺苷酸化
chickenpreadipocytes3'-UTRalternative polyadenylation(APA)
《畜牧兽医学报》 2026 (6)
3157-3169,13
湖北省重大攻关项目(2023BAA029)
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