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非综合征性口腔颌面部裂隙中的长链非编码RNA MALAT1和NEAT1

Long non-coding RNAs MALAT1 and NEAT1 in non-syndromic orofacial clefts.

作者信息

Wang Errui, Guo Yumeng, Gao Shuting, Zhou Ying, Liu Bin, Dissanayaka Waruna Lakmal, Zheng Yayuan, Zhou Qiaozhen, Zhai Junkai, Gao Zhengkun, Zhang Baoping, Liu Ruimin, Zhang Kailiang

机构信息

School of Stomatology, Lanzhou University, Lanzhou, China.

Applied Oral Sciences & Community Dental Care, Faculty of Dentistry, The University of Hong Kong, Hong Kong, China.

出版信息

Oral Dis. 2023 May;29(4):1668-1679. doi: 10.1111/odi.14177. Epub 2022 Mar 16.

Abstract

Long non-coding RNAs (lncRNAs) are thought to play important roles in non-syndromic orofacial clefts (NSOFC). Clinical diagnosis was categorized as either non-syndromic cleft lip with or without cleft palate (NSCL/P), or non-syndromic cleft palate only (NSCPO). Tissues excised from the trimmed wound edge were reserved as experimental samples; adjacent normal control was used as a positive control, and tissue from healthy individuals was used as a blank control. Target lncRNAs in the collected tissues were identified using microarrays and quantitative reverse transcription PCR (RT-qPCR). Immunohistochemical (IHC) staining and RT-qPCR were used to verify the target mRNAs. Pathway, gene ontology (GO) enrichment, and TargetScan predictions were employed to construct competing endogenous RNA networks (ceRNA networks) and explore their potential functions. RNA-Seq revealed 24 upregulated and 43 downregulated lncRNAs; MALAT1 and NEAT1 were screened and validated using RT-qPCR. Common NSOFC risk factors were positively correlated with MALAT1 and NEAT1 expression. Bioinformatics predicted four ceRNA networks; GO enrichment focused on their potential functions. RT-qPCR and IHC data were consistent with respect to expression levels of proteins and the mRNAs that encode them. As MALAT1 and NEAT1 are associated with the severity of NSOFC, they represent potential therapeutic targets and prognostic biomarkers.

摘要

长链非编码RNA(lncRNAs)被认为在非综合征性口腔颌面部裂隙(NSOFC)中发挥重要作用。临床诊断分为非综合征性唇裂伴或不伴腭裂(NSCL/P),或仅非综合征性腭裂(NSCPO)。从修剪后的伤口边缘切除的组织留作实验样本;相邻的正常对照用作阳性对照,健康个体的组织用作空白对照。使用微阵列和定量逆转录PCR(RT-qPCR)鉴定收集组织中的靶lncRNAs。免疫组织化学(IHC)染色和RT-qPCR用于验证靶mRNA。采用通路、基因本体(GO)富集和TargetScan预测构建竞争性内源性RNA网络(ceRNA网络)并探索其潜在功能。RNA测序揭示了24个上调和43个下调的lncRNAs;使用RT-qPCR筛选并验证了MALAT1和NEAT1。常见的NSOFC危险因素与MALAT1和NEAT1表达呈正相关。生物信息学预测了四个ceRNA网络;GO富集关注其潜在功能。RT-qPCR和IHC数据在蛋白质及其编码mRNA的表达水平方面是一致的。由于MALAT1和NEAT1与NSOFC的严重程度相关,它们代表潜在的治疗靶点和预后生物标志物。

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