摘要：

为了探究lncRNA2099对猪前脂肪细胞增殖分化的影响,采用实时荧光定量PCR(qRT-PCR)技术检测lncRNA2099在猪心脏、肝脏、脾脏、肺脏、肾脏、背肌和背脂及离体培养的前体脂肪细胞增殖和分化阶段的表达水平;利用核质分离技术确定lncRNA2099在脂肪细胞中的表达位置;构建真核表达载体pcDNA3.1-lncRNA2099,采用qRT-PCR技术检测在脂肪细胞内过表达该lncRNA后对脂肪细胞增殖、分化相关标志基因表达的影响。结果显示:lncRNA2099存在明显组织表达特异性,且在肾脏和背脂中高表达,在背肌中不表达;lncRNA2099在猪前脂肪细胞增殖阶段12 h表达量最高,随后逐渐降低。在分化阶段第2 天表达量最高,随后逐渐降低。核质定位结果表明,lncRNA2099在细胞核与细胞质中均有表达;在脂肪细胞内过表达lncRNA2099后,增殖标志基因P21与成脂分化标志基因LPL表达量极显著升高。lncRNA2099可能作为转录调节因子抑制猪前脂肪细胞增殖、促进成脂分化。

关键词: 猪, lncRNA2099, 前脂肪细胞, 增殖, 成脂分化

Abstract:

To investigate the effect of lncRNA2099 on the proliferation and differentiation of porcine preadipocytes,the qRT-PCR was used to detect the expression level of lncRNA2099 in heart,liver,spleen,lung,kidney,back muscle,back fat and proliferation and differentiation stages of porcine preadipocyte.The nucleoplasmic separation technique was used to determine the expression location of lncRNA2099 in adipocyte.The eukaryotic expression vector pcDNA3.1-lncRNA2099 was constructed and qRT-PCR was used to detect its impact on the expression of marker genes related to adipocyte proliferation and differentiation after over expression in adipocyte.The results showed that lncRNA2099 was highly expressed in kidney and back fat,but not in back muscle.The nucleoplasmic localization results showed that lncRNA2099 was expressed in both nucleus and cytoplasm;the expression levels of proliferation marker gene P21 and lipogenic differentiation marker gene LPL significantly increased after overexpression of lncRNA2099 in adipocytes.lncRNA2099 may act as a transcriptional regulator to inhibit proliferation and promote lipogenic differentiation in porcine preadipocytes.

Key words: Pig, lncRNA2099, Preadipocytes, Proliferation, Lipogenic differentiation