山东医药2018，Vol.58Issue(8)：1-5,5.DOI:10.3969/j.issn.1002-266X.2018.08.001
短扩增子对结直肠癌转移石蜡组织LncRNA定量分析的可行性
Feasibility of quantitative analysis of lncRNA in FFPE tissues of metastatic colorectal cancer with short amplicons
摘要
Abstract
Objective To explore the feasibility of quantitative analysis of long non-coding RNA (lncRNA) in FFPE tissues of metastatic colorectal cancer with short amplicons. Methods We selected 4 target lncRNAs associated with colorectal cancer metastasis (LncRNA-Enst00000554679, LncRNA-Enst00000550851, LncRNA-Enst00000497498, and LncRNA-Enst00000513016). Fifty-five cases of FFPE tissue specimens and 40 cases of frozen tissue specimens were collected, each specimen contained the colorectal cancer tissues, adjacent normal tissues, and lymph node metastasis tissues. The qRT-PCR was used to detect the expression of 4 LncRNAs after amplification of long amplicons in 25 samples of colorectal cancer tissues, adjacent normal tissues and metastatic lymph node tissues. Four lncRNA short amplicon primers were designed to assess the amplification efficiency and the assay coefficient (R2) of long and short amplicons in frozen and FFPE tissues. The quantitative concordance of short amplicons and long amplicons in the frozen tissues was analyzed, as well as the quantitative concordance of short amplicons in the frozen tissues and FFPE tissues. We separately designed three pairs of short amplicon primers for four lncRNAs, and analyzed the quantitative concordance of three short amplicons in the FFPE tissues. Results Four lncRNAs expressed significantly higher in the colorectal cancer tissues (all P＜0. 05). Short amplicons could achieve the best amplification efficiency in both frozen and FFPE tissues, while the long amplicons did not reach the best amplification efficiency. The R2 of the short amplicon was significantly higher than that of the long amplicon(P＜0. 05). The quantitative concordance of short and long amplicons in the frozen tissues was 62. 5％, and that of short amplicons in the frozen and FFPE tissues was only 25. 0％. The coincidence rate of the three short amplicons in the four LncRNAs was 37. 5％, while that of the two short amplicons was 100％. Conclusion Four lncRNAs are highly expressed in the metastatic colorectal cancer tissues, and the short amplicons can be used for lncRNA quantitative analysis in the FFPE tissues.关键词
结直肠癌/肿瘤转移/长链非编码RNA/石蜡组织/实时荧光定量PCRKey words
colorectal carcinoma/neoplasm metastasis/long non-coding RNA/paraffin-embedded tissue/real-time fluorescent quantitative PCR分类
医药卫生引用本文复制引用
李佳茜,孙青..短扩增子对结直肠癌转移石蜡组织LncRNA定量分析的可行性[J].山东医药,2018,58(8):1-5,5.基金项目
国家自然科学基金资助项目(81272420) （81272420）
山东省自然科学基金资助项目(ZR2015HL066). （ZR2015HL066）
