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人牙周膜细胞在张力加载下的 mRNA 和长非编码 RNA 表达谱分析。

mRNA and long non-coding RNA expression profiling of human periodontal ligament cells under tension loading.

机构信息

Division of Paediatric Dentistry and Orthodontics, Faculty of Dentistry, The University of Hong Kong, Hong Kong SAR, China.

Division of Periodontology and Implant Dentistry, Faculty of Dentistry, The University of Hong Kong, Hong Kong SAR, China.

出版信息

Eur J Orthod. 2021 Dec 1;43(6):698-707. doi: 10.1093/ejo/cjab043.

Abstract

OBJECTIVE

This study explored the expression profiles of messenger RNAs (mRNAs) and long non-coding RNAs (lncRNAs) in human periodontal ligament (PDL) cells subjected to tensile loading.

METHODS

PDL cells were isolated from the teeth of five healthy individuals, cultured and then exposed to tensile loading. RNA sequencing was performed to explore the mRNA and lncRNA expression profiles with or without tensile loading. Differential expression, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted to reveal enriched biological functions and signal transduction pathways. Quantitative polymerase chain reaction (qPCR) was performed to validate the expression of specific mRNAs and lncRNAs associated with the enriched pathways.

RESULTS

Tensile loading significantly enhanced the osteogenic potential of PDL cells. Overall, 1438 mRNAs (860 up- and 578 down-regulated) and 195 lncRNAs (107 up- and 88 down-regulated) were differentially expressed (adjusted P-value <0.05) in the tensile loading group versus the control group. GO and KEGG analyses of the differentially expressed genes indicated significant enrichment in osteogenesis-related biological processes and intracellular signal transduction pathways (e.g. the PI3K-Akt pathway), respectively. The qPCR analysis validated the expression levels of five selected mRNAs (EGFR, FGF5, VEGFA, HIF1A, and FOXO1) and three selected lncRNAs (CYTOR, MIR22HG, and SNHG3).

LIMITATION

Further studies are warranted to validate the mechanisms regulating tension-induced bone remodelling in PDL cells and potential regulation by the identified lncRNAs.

CONCLUSION

The notably altered mRNA and lncRNA expression profiles in PDL cells under tensile loading enhance our mechanistic understanding of tension-induced osteogenesis.

摘要

目的

本研究探讨了在受到拉伸载荷的情况下,人牙周韧带(PDL)细胞中信使 RNA(mRNA)和长链非编码 RNA(lncRNA)的表达谱。

方法

从五名健康个体的牙齿中分离出 PDL 细胞,进行培养,然后暴露于拉伸载荷下。进行 RNA 测序以探讨有无拉伸载荷时的 mRNA 和 lncRNA 表达谱。进行差异表达、基因本体论(GO)和京都基因与基因组百科全书(KEGG)富集分析,以揭示富集的生物学功能和信号转导途径。采用实时定量聚合酶链反应(qPCR)验证与富集途径相关的特定 mRNAs 和 lncRNAs 的表达。

结果

拉伸载荷显著增强了 PDL 细胞的成骨潜能。总体而言,在拉伸载荷组与对照组相比,有 1438 个 mRNAs(860 个上调和 578 个下调)和 195 个 lncRNAs(107 个上调和 88 个下调)差异表达(调整后的 P 值<0.05)。差异表达基因的 GO 和 KEGG 分析分别表明,在成骨相关的生物学过程和细胞内信号转导途径(如 PI3K-Akt 途径)中存在显著富集。qPCR 分析验证了五个选定的 mRNAs(EGFR、FGF5、VEGFA、HIF1A 和 FOXO1)和三个选定的 lncRNAs(CYTOR、MIR22HG 和 SNHG3)的表达水平。

局限性

需要进一步的研究来验证调节 PDL 细胞中张力诱导骨重塑的机制以及鉴定的 lncRNAs 的潜在调节作用。

结论

在拉伸载荷下,PDL 细胞中明显改变的 mRNA 和 lncRNA 表达谱增强了我们对张力诱导成骨的机制理解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/af66/8643418/ab8f79d3a2ed/cjab043f0001.jpg

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