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光老化皮肤中 LncSPRY4-IT1 靶向功能蛋白的鉴定和生物学分析。

Identification and biological analysis of LncSPRY4-IT1-targeted functional proteins in photoaging skin.

机构信息

Department of Dermato-venereology, The Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China.

Department of Dermato-venereology, The Third Affiliated Hospital of Sun Yat-sen University -Yuedong Hospital, Meizhou, China.

出版信息

Photodermatol Photoimmunol Photomed. 2021 Nov;37(6):530-538. doi: 10.1111/phpp.12705. Epub 2021 Jun 26.

Abstract

BACKGROUND/PURPOSE: Skin photoaging, main causes of skin aging, is induced by chronic UV irradiation. LncSPRY4-IT1, a broadly expressed lncRNA, takes part in various biological functions by combining with functional protein molecules. However, the role of LncSPRY4-IT1 in skin photoaging process has not been characterized. This study is to investigate the interacting proteins of LncSPRY4-IT1 by combining RNA pull-down, high-throughput, and bioinformatic analysis.

METHODS

Human skin fibroblasts (HDFs) were exposed to 10 J/cm UVA irradiation, once a day for 14 days. LncSPRY4-IT1 expression was qualified via RT-PCR. In vitro RNA pull-down assays and liquid chromatography-mass spectrometry analysis were used to identify the LncSPRY4-IT1-related proteins. Functional annotation analysis and pathway enrichment were preformed via Gene Ontology and KEGG.

RESULTS

LncSPRY4-IT1 expression in photoaging fibroblasts was increased 1.66 ± 0.23 folds. 181 LncSPRY4-IT1-interacting proteins in UVA-induced photoaging skin fibroblast irradiation were identified, of which 56 proteins with two or more unique peptides, 73 proteins related to RNA processing, and 5 proteins related to DNA processing. High-throughput and bioinformatic analysis showed that LncSPRY4-IT1-targeting proteins were involved in cellular process, metabolic process, biological regulation, and cell part in skin photoaging process. The KEGG revealed that LncSPRY4-IT1-targeting proteins were mainly enriched in metabolic pathways.

CONCLUSION

The results of our studies illuminate how LncSPRY4-IT1 formed a LncRNA-protein regulatory network in skin photoaging mechanisms and suggest that LncSPRY4-IT1 may serve as a novel upstream intervention target for the prevention and treatment of photoaging and related skin diseases.

摘要

背景/目的:皮肤光老化是皮肤衰老的主要原因,由慢性紫外线照射引起。广泛表达的长链非编码 RNA LncSPRY4-IT1 通过与功能蛋白分子结合参与多种生物学功能。然而,LncSPRY4-IT1 在皮肤光老化过程中的作用尚未得到描述。本研究通过 RNA 下拉、高通量和生物信息学分析,结合研究 LncSPRY4-IT1 的相互作用蛋白。

方法

将人皮肤成纤维细胞(HDF)暴露于 10 J/cm UVA 照射下,每天一次,共 14 天。通过 RT-PCR 鉴定 LncSPRY4-IT1 的表达。采用体外 RNA 下拉实验和液相色谱-质谱分析鉴定 LncSPRY4-IT1 相关蛋白。通过基因本体论和京都基因与基因组百科全书进行功能注释分析和通路富集分析。

结果

光老化成纤维细胞中 LncSPRY4-IT1 的表达增加了 1.66±0.23 倍。在 UVA 诱导的光老化皮肤成纤维细胞照射中鉴定出 181 个 LncSPRY4-IT1 相互作用蛋白,其中 56 个蛋白具有两个或更多独特肽段,73 个蛋白与 RNA 处理有关,5 个蛋白与 DNA 处理有关。高通量和生物信息学分析表明,LncSPRY4-IT1 靶向蛋白参与皮肤光老化过程中的细胞过程、代谢过程、生物调节和细胞部分。KEGG 显示 LncSPRY4-IT1 靶向蛋白主要富集在代谢途径中。

结论

本研究结果阐明了 LncSPRY4-IT1 在皮肤光老化机制中形成 LncRNA-蛋白调控网络的方式,并表明 LncSPRY4-IT1 可能作为皮肤光老化及相关皮肤疾病防治的新型上游干预靶点。

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