文献类型：期刊文献

中文题名：持续静压力环境下MC3T3-E1细胞的基因测序分析

英文题名：Gene sequencing analysis of MC3T3-E1 cells under continuously compressive pressures

作者：宋志靖[1];张浩令[2];蒋宇航[2];王凯[1];宁浩驰[1];裴雪冬[1];陈海亮[1];宋敏[1];王薇[3]

第一作者：宋志靖

机构：[1]甘肃中医药大学中医临床学院,兰州730000;[2]甘肃中医药大学公共卫生学院,兰州730000;[3]甘肃中医药大学针灸推拿学院,兰州730000

第一机构：甘肃中医药大学中医临床学院

年份：2022

卷号：47

期号：5

起止页码：529

中文期刊名：重庆医科大学学报

外文期刊名：Journal of Chongqing Medical University

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD_E2021_2022】；

基金：国家自然科学基金资助项目(编号:81960877)

语种：中文

中文关键词：MC3T3-E1细胞;持续静压力;基因测序;骨质疏松症

外文关键词：MC3T3-E1 cell;continuously compressive pressure;gene sequencing;osteoporosis

摘要：目的:通过对持续静压力环境下MC3T3-E1细胞的基因测序,筛选出Hub基因并分析其与骨质疏松症相关性。方法:采用MTT检测细胞增殖和免疫荧光检测细胞骨架蛋白,筛选出最佳实验条件。用TRIzol法提取RNA与基因测序,通过Mfuzz聚类、富集分析、GSVA分析,利用NCBI GEO数据库、Genecards数据库、molecular signatures database数据库,分析对比持续静压力环境下MC3T3-E1细胞差异表达基因,筛选Hub基因,对比分析其与骨质疏松症信号通路关联的分子学机制。结果:在持续静压力环境下,MC3T3-E1细胞增殖显著降低,在1.0 MPa处理8 h和0.5 MPa处理24 h细胞模型组的细胞骨架中,微管蛋白和肌动蛋白的抑制作用显著。通过空白组与模型组基因测序对比后发现29164个差异基因,其中14489个上调、14675个下调。这些差异基因涉及1658个GO功能,包括1096 GO_BP(生物过程),255 GO_CC(细胞组分),307 GO_MF(分子功能),MC3T3-E1细胞在持续静压力作用下受到差异基因调控的信号转导通路有305个。最后筛选出的Hub基因为BTK、CSF1R、MATK、NOS1、PDGFRB,骨质疏松症中MATK表达显著(P<0.05),与5个核心基因相对应的AUC曲线下面积为BTK(0.900)、CSF1R(0.850)、MATK(0.950)、NOS1(0.650)、PDGFRB(0.800)。通过对5个核心基因进行GSVA分析,得出MATK涉及3个关键信号传导途径:INFLAMMATORY-RESPONSE、HEDGEHOG-SIGNALING和KRAS-SIGNALING-DN。结论:持续静压力可降低MC3T3-E1细胞增殖活性,并降低细胞骨架中微管蛋白和肌动蛋白表达;MATK可能作为持续静压力环境下调控MC3T3-E1细胞成骨基因,并与骨质疏松症3个关键信号传导途径相关。
Objective:To screen out the Hub gene and analyze its correlation with osteoporosis by sequencing the genes of MC3T3-E1 cells under continuously compressive pressures(CCP).Methods:MTT assay was used to detect cell proliferation and immunofluorescence assay was used to screen out the best experimental conditions.RNA was extracted by TRIzol method and sequenced.And by Mfuzz clustering,enrichment analysis,GSVA analysis,NCBI GEO database,Genecards database and molecular signatures database,the differentially expressed genes in MC3T3-E1 cells under CCP were analyzed and compared,the Hub gene was screened,and the molecular mechanism associated with osteoporosis signal pathway was compared and analyzed.Results:The proliferation of MC3T3-E1 cells decreased significantly under CCP,and tubulin and actin inhibited significantly in the cytoskeleton of cell model groups treated with 1.0 MPa for 8 hours and 0.5 MPa for 24 hours.After comparing the blank group and the model group,29164 different genes were found,of which 14489 were up-regulated and 14675 were down-regulated.These differential genes involved 1658 GO functions,including 1096 GO_BP(biological process),255 GO_CC(cell component)and 307 GO_MF(molecular function),and there were 305 signal transduction pathways regulated by differential genes in MC3T3-E1 cells under CCP.The final screened Hub genes were BTK,CSF1R,MATK,NOS1 and PDGFRB.The expression of MATK in osteoporosis was significant(P<0.05).The areas under AUC curve corresponding to the five core genes were BTK(0.900),CSF1R(0.850),MATK(0.950),NOS1(0.650)and PDGFRB(0.800).According to GSVA analysis of five core genes,MATK was involved in three key signaling pathways:INFLAMMATORY-RESPONSE,HEDGEHOG-SIGNALING and KRAS-SIGNALING-DN.Conclusion:CCP can decrease the proliferation activity of MC3T3-E1 cells,and reduce the influence of tubulin and actin in cytoskeleton.MATK may be a gene that regulates the osteogenesis of MC3T3-E1 cells under CCP,and is related to three key signal transmission pathways of osteoporosis.