基于转录组测序筛选鹿茸胸腺素促成骨细胞增殖基因  

Screening of genes that promote osteoblast proliferation by thymosinβ4 from sika antler based on transcriptome sequencing

作　　者：薛东明 慧芳 孙天霞 姜英男 赵雨[1] XUE Dongming;HUI Fang;SUN Tianxia;JIANG Yingnan;ZHAO Yu(Jilin Ginseng Academy,Changchun University of Chinese Medicine,Changchun 130117,China)

机构地区：[1]长春中医药大学人参科学研究院,吉林长春130117

出　　处：《中国骨质疏松杂志》2023年第1期17-22,共6页Chinese Journal of Osteoporosis

基　　金：国家科技部重点研发计划(2018YFC1706603)。

摘　　要：目的对Tβ4蛋白处理组和空白组进行转录组测序,筛选Tβ4蛋白处理后促成骨细胞相关的候选基因和通路。方法通过Fmoc固相合成法体外合成梅花鹿Tβ4蛋白,并处理MC3T3细胞,CCK8法检测细胞活性。将细胞分为空白组和Tβ4处理组,利用Illumina平台进行高通量测序。对差异表达基因进行GO功能富集分类和KEGG通路分析,构建差异基因的蛋白互作网络并筛选Hub基因,通过在线软件对Hub基因进行富集分析。结果Tβ4蛋白处理成骨细胞显著促进了细胞的增殖活性。获得差异基因6106个,其中上调基因1979个,下调基因4127个。根据PPI网络中的节点连通度,得到10个Hub基因,其中,筛选到7个与成骨细胞增殖相关的候选基因,分别为CDC20、PLK1、CDK1、BUB1B、BUB1、MAD2L1、RRM2,这些基因富集到了细胞周期信号通路和p53信号通路。结论梅花鹿Tβ4蛋白处理后可以提高细胞的增殖活性和存活率。梅花鹿胸腺素Tβ4可能是通过调控细胞周期信号通路和p53信号通路中的关键基因达到促进成骨细胞增殖的功能。Objective To screen the candidate genes and pathways related to osteoblast after Tβ4 protein treatment by transcriptome sequencing of Tβ4 protein treated group and blank group.Methods The sika deer Tβ4 protein was synthesized using Fmoc solid-phase synthesis in vitro and applied to MC3T3 cells.The cell activity was detected with CCK8 method.The cells were divided into blank group and Tβ4-treated group.The high-throughput sequencing was carried out with Illumina platform.The differently expressed genes were analyzed with GO functional enrichment classification and KEGG pathway analysis.The protein interaction network of differential genes was constructed and Hub genes were screened.Hub genes were enriched and analyzed using online software.Results The cell proliferation was significantly promoted in osteoblasts treated with Tβ4 protein.A total of 6,106 differential genes were obtained,including 1979 up-regulated genes and 4127 down-regulated genes.According to the node connectivity of PPI network,10 Hub genes were obtained,among which 7 candidate genes related to osteoblast proliferation were screened.They were CDC20,PLK1,CDK1,BUB1B,BUB1,MAD2L1,and RRM2,respectively.These genes were enriched in cell cycle signal pathway and p53 signal pathway.Conclusion Sika deer Tβ4 protein treatment improves the cell proliferation activity and survival rate.Thymosinβ4 in sika deer may promote osteoblast proliferation by regulating key genes in cell cycle signal pathway and p53 signal pathway.

关 键 词：鹿茸 胸腺素Β4 转录组测序 细胞增殖 成骨细胞 

分 类 号：R932[医药卫生—生药学]