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口腔鳞状细胞癌福尔马林固定石蜡包埋组织中长链非编码RNA的定量分析

Quantitative analysis of lncRNA in formalin-fixed paraffin-embedded tissues of oral squamous cell carcinoma.

作者信息

Kumar Kiran, Hallikeri Kaveri, Oli Ajaykumar, Goni Mallikarjun, Jain Apoorva, Poyya Jagadeesha, Shilpasree Alagilavada S, Javaregowda Palaksha Kanive

机构信息

Department of Oral and Maxillofacial Pathology and Oral Microbiology, SDM College of Dental Sciences and Hospital (a constituent unit of Shri Dharmasthala Manjunatheshwara University), Dharwad, Karnataka State, 580009, India.

Department of Biomedical Science, SDM Research Institute for Biomedical Sciences (a constituent unit of Shri Dharmasthala Manjunatheshwara University), Dharwad, Karnataka State, 580009, India.

出版信息

Biotechniques. 2023 Oct;75(4):133-142. doi: 10.2144/btn-2023-0033. Epub 2023 Aug 17.

Abstract

The study evaluated expression profiles of few regulatory lncRNAs in oral squamous cell carcinoma and normal mucosa adjacent to oral cancer using paired fresh frozen and formalin-fixed paraffin-embedded (FFPE) tissues stored at a different duration of time (1-5 years) using real-time quantitative PCR. The quantity and quality of total RNA isolated from FFPE tissues was less compared with that of fresh frozen tissues, which resulted in a noncorrelation of quantification cycle values. Following normalization, the expression of lncRNAs in the paired tissues did not differ significantly. The differential expression of the lncRNAs in the study was consistent with The Cancer Genome Atlas head and neck squamous cell carcinoma database. The study findings demonstrate the possibility of performing accurate quantitative analysis of lncRNAs using short amplicons and standardized real-time quantitative PCR assays in oral squamous cell carcinoma FFPE samples.

摘要

本研究使用实时定量PCR,对存储不同时长(1 - 5年)的配对新鲜冰冻组织和福尔马林固定石蜡包埋(FFPE)组织,评估了口腔鳞状细胞癌及口腔癌旁正常黏膜中少数调控性长链非编码RNA(lncRNA)的表达谱。与新鲜冰冻组织相比,从FFPE组织中分离的总RNA的数量和质量较低,这导致定量循环值不相关。标准化后,配对组织中lncRNA的表达无显著差异。本研究中lncRNA的差异表达与癌症基因组图谱头颈鳞状细胞癌数据库一致。研究结果表明,在口腔鳞状细胞癌FFPE样本中,使用短扩增子和标准化实时定量PCR检测对lncRNA进行准确定量分析是可行的。

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