永生化麝鼠香腺成纤维细胞的转录组学分析

Transcriptome Analysis of Immortalized Muskrat Scent Gland Fibroblasts

【目的】探索永生化对麝鼠香腺成纤维细胞转录组的影响。【方法】在获得永生化的麝鼠香腺成纤维细胞后,采用二代测序的方法对其永生化前后转录组进行检测,并通过生物信息学方法进行差异分析。【结果】用Trinity软件对麝鼠永生化和原代香腺细胞测序数据进行基因组装,总共发现62 152 unigenes,富集分析结果显示,上述基因主要聚集在细胞代谢、细胞生物学调控、蛋白结合和酶催化等基因的细胞基础代谢活动。基于DESeq2包的基因表达差异分析结果表明,永生化和原代细胞表达差异的基因总共有8 690个,其中上调基因3 414个,下调基因5 276个,上调基因主要富集于DNA复制、同源重组、错配修复、细胞周期和RNA代谢通路,下调基因主要富集于氧化磷酸化、帕金森病和阿尔茨海默病疾病通路中。【结论】永生化过程对麝鼠原代细胞基因的影响较大,可为后续建立永生化香腺细胞体外泌香体系提供理论支撑。

【Objective】To explore the influence of immortalization on transcriptome of primary muskrat scent gland fibroblasts.【Method】Muskrat gland cells are the key to secreting muskrat musk. After obtaining the immortalized muskrat gland fibroblasts,in this study,transcriptome of muskrat immortalized and primary cells were detected through second-generation sequencing method,and the differences were analyzed by bioinformatics method.【Result】A total of 62 152 unigenes were found by gene assembling of the sequencing data of immortalized and primary cells using Trinity software. Cluster analysis showed that these genes were mainly enriched in cell metabolism,cell biological regulation,protein binding,enzyme catalysis and other basic metabolic activities. A total of 8 690 differentially expressed genes were detected according to DESeq2 analysis results,including 3 414 up-regulated and 5 276 downregulated genes. And the up-regulated genesmainly occurred in DNA replication,raised homologous recombination,mismatch repair,cell cycle and RNA metabolism pathway,while down-regulated genes mainly enriched in the oxidative phosphorylation,Parkinson’s and Alzheimer moss disease pathways in the disease.【Conclusion】Immortalization process had great influence on muskrat primary scent gland genes. This study provided theoretical support for the subsequent establishment of musk secretion system of immortalized scent glands cells in vitro.