RNA‐sequencing analysis reveals the potential molecular mechanism of RAD54B in the proliferation of inflamed human dental pulp cells

小桶 小干扰RNA 免疫印迹 生物 分子生物学 流式细胞术 核糖核酸 基因 基因表达 细胞周期 转染 细胞生物学 转录组 遗传学
作者
Huang Pei,Fushi Wang,Xinhuan Wang,Xiujiao Meng,Weiwei Qiao,Liuyan Meng
出处
期刊:International Endodontic Journal [Wiley]
卷期号:56 (1): 39-52 被引量:1
标识
DOI:10.1111/iej.13842
摘要

To investigate the role of RAD54B in the proliferation of inflamed human dental pulp cells (hDPCs) induced by lipopolysaccharide (LPS).Normal, carious and pulpitic human dental pulp tissues were collected. Total RNA was subjected to RNA-sequencing (seq) and gene expression profiles were studied by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Differentially expressed genes (DEGs) in homologous recombination repair (HRR) were validated with qRT-PCR. The expression of RAD54B and TNF-α in human dental pulp tissues was detected using immunohistochemistry. HDPCs were cultured and RAD54B level in hDPCs was detected after LPS stimulation using western blot. CCK-8 was used to investigate the proliferation of hDPCs transfected with negative control (Nc) small interfering RNA (siRNA), RAD54B siRNA, P53 siRNA or both siRNAs with or without LPS stimulation. Flow cytometry was used to detect the cell cycle distribution, and western blot and immunofluorescence were used to analyse the expression of RAD54B, P53 and P21 under the above treatments. One-way and two-way anova followed by least significant difference posttest were used for statistical analysis.RNA-seq results identified DEGs amongst the three groups. KEGG pathway analysis revealed enrichment of DEGs in the replication and repair pathway. HRR and non-homologous end joining (NHEJ) components were further verified and qRT-PCR results were basically consistent with the sequencing data. RAD54B, an HRR accessory factor highly expressed in carious and pulpitic tissues as compared to that in normal pulps, was chosen as our gene of interest. High RAD54B expression was confirmed in inflamed human dental pulp tissues and LPS-stimulated hDPCs. Upon RAD54B knockdown, P53 and P21 expressions in hDPCs were upregulated whereas the proliferation was significantly downregulated, accompanied by increased G2/M phase arrest. After inhibiting P53 expression in RAD54B-knockdown hDPCs, P21 expression and cell proliferation were reversed.Gene expression profiles of normal, carious and pulpitic human dental pulp tissues were revealed. HRR components were elucidated to function in dental pulp inflammation. Amongst the DEGs in HRR, RAD54B regulated the proliferation of inflamed hDPCs via P53/P21 signalling. This research deepens our understanding of dental pulp inflammation and provides new insight to clarify the underlying mechanisms.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
刚刚
2秒前
尊敬的发夹完成签到,获得积分10
2秒前
陆陆发布了新的文献求助10
3秒前
Lucas应助包容的幻梅采纳,获得10
3秒前
团团团完成签到 ,获得积分10
3秒前
渝州人完成签到,获得积分10
3秒前
su发布了新的文献求助10
3秒前
生动大白菜真实的钥匙完成签到,获得积分10
4秒前
Asystasia7发布了新的文献求助10
4秒前
4秒前
萧然完成签到,获得积分10
5秒前
鲤鱼听荷完成签到 ,获得积分10
5秒前
虚心梦秋发布了新的文献求助10
6秒前
莱昂纳多的李完成签到,获得积分10
6秒前
开心烨磊完成签到,获得积分20
6秒前
6秒前
三里墩头完成签到,获得积分10
6秒前
娃哈哈完成签到 ,获得积分10
7秒前
7秒前
Jasper应助老六采纳,获得10
7秒前
lz完成签到,获得积分10
7秒前
Estrella完成签到,获得积分10
7秒前
ZLX完成签到,获得积分10
7秒前
XY打钉佬完成签到 ,获得积分10
8秒前
大个应助陈粮酿好酒采纳,获得10
8秒前
anthea完成签到 ,获得积分10
8秒前
北冰石完成签到,获得积分10
8秒前
呆萌棒棒糖完成签到,获得积分10
8秒前
sevten完成签到,获得积分10
9秒前
bkagyin应助钮祜禄则天采纳,获得10
9秒前
柠檬九分酸完成签到,获得积分10
9秒前
飞鱼发布了新的文献求助10
9秒前
szy完成签到,获得积分10
9秒前
10秒前
Y.J完成签到,获得积分10
10秒前
thangxtz完成签到,获得积分10
10秒前
snowpaper完成签到,获得积分10
10秒前
dl发布了新的文献求助10
10秒前
MrX完成签到,获得积分10
10秒前
高分求助中
All the Birds of the World 4000
Production Logging: Theoretical and Interpretive Elements 3000
Les Mantodea de Guyane Insecta, Polyneoptera 2000
Machine Learning Methods in Geoscience 1000
Resilience of a Nation: A History of the Military in Rwanda 888
Essentials of Performance Analysis in Sport 500
Measure Mean Linear Intercept 500
热门求助领域 (近24小时)
化学 材料科学 医学 生物 工程类 有机化学 物理 生物化学 纳米技术 计算机科学 化学工程 内科学 复合材料 物理化学 电极 遗传学 量子力学 基因 冶金 催化作用
热门帖子
关注 科研通微信公众号,转发送积分 3729470
求助须知:如何正确求助?哪些是违规求助? 3274545
关于积分的说明 9986446
捐赠科研通 2989686
什么是DOI,文献DOI怎么找? 1640734
邀请新用户注册赠送积分活动 779332
科研通“疑难数据库(出版商)”最低求助积分说明 748188