Gene sequencing analysis of MC3T3-E1 cells under continuously compressive pressures

Song Zhijing; Zhang Haoling; Jiang Yuhang; Wang Kai; Ning Haochi; Pei Xuedong; Chen Hailiang; Song Min; Wang Wei

doi:10.13406/j.cnki.cyxb.002851

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Gene sequencing analysis of MC3T3-E1 cells under continuously compressive pressures

更新时间：2021-10-08

- Gene sequencing analysis of MC3T3-E1 cells under continuously compressive pressures
- Journal of Chongqing Medical University (2021)
- 作者机构：
  
  1. 甘肃中医药大学中医临床学院
  2. 甘肃中医药大学公共卫生学院
  3. 甘肃中医药大学针灸推拿学院
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.002851
  CLC： R580
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[1]宋志靖,张浩令,蒋宇航,王凯,宁浩驰,裴雪冬,陈海亮,宋敏,王薇.持续静压力环境下MC3T3-E1细胞的基因测序分析[J/OL].重庆医科大学学报:1-9[2021-06-23].https://doi.org/10.13406/j.cnki.cyxb.002851.

Song Zhijing, Zhang Haoling, Jiang Yuhang, et al. Gene sequencing analysis of MC3T3-E1 cells under continuously compressive pressures. [J/OL]. Journal of Chongqing Medical University (2021)
[1]宋志靖,张浩令,蒋宇航,王凯,宁浩驰,裴雪冬,陈海亮,宋敏,王薇.持续静压力环境下MC3T3-E1细胞的基因测序分析[J/OL].重庆医科大学学报:1-9[2021-06-23].https://doi.org/10.13406/j.cnki.cyxb.002851. DOI：

Song Zhijing, Zhang Haoling, Jiang Yuhang, et al. Gene sequencing analysis of MC3T3-E1 cells under continuously compressive pressures. [J/OL]. Journal of Chongqing Medical University (2021) DOI： 10.13406/j.cnki.cyxb.002851.

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29 164个差异基因

其中14 489个上调、14 675个下调。这些差异基因涉及1 658个GO功能

包括1 096 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。

Abstract

Objective:To screen out the Hub gene and analyze its correlation with osteoporosis by sequencing the genes of MC3 T3-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 MC3 T3-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 MC3 T3-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

29 164 different genes were found

of which 14 489 were up-regulated and 14 675 were down-regulated.These differential genes involve 1 658 GO functions

including 1 096 GO BP（biological process）

255 GO CC（cell component） and 307 GO MF（molecular function）

and there are 305 signal transduction pathways regulated by differential genes in MC3 T3-E1 cells under CCP.The final screened Hub genes are BTK

CSF1 R

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）

CSF1 R（0.850）

MATK（0.950）

OS1（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 MC3 T3-E1 cells

and reduce the influence of tubulin and actin in cytoskeleton.MATK may be a gene that regulates the osteogenesis of MC3 T3-E1 cells under CCP

and is related to three key signal transmission pathways of osteoporosis.

关键词

MC3T3-E1细胞持续静压力基因测序骨质疏松症

Keywords

MC3T3-E1 cellscontinuously compressive pressuresgene sequencingosteoporosis

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