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通过nCounter技术鉴定出ACTB、CDKN1B、GAPDH、GRB2、RHOA和SDCBP作为神经内分泌肺癌中的参考基因。

ACTB, CDKN1B, GAPDH, GRB2, RHOA and SDCBP Were Identified as Reference Genes in Neuroendocrine Lung Cancer via the nCounter Technology.

作者信息

Walter Robert Fred Henry, Werner Robert, Vollbrecht Claudia, Hager Thomas, Flom Elena, Christoph Daniel Christian, Schmeller Jan, Schmid Kurt Werner, Wohlschlaeger Jeremias, Mairinger Fabian Dominik

机构信息

Ruhrlandklinik, West German Lung Center, University Hospital Essen, University of Duisburg-Essen, Essen, Germany.

Institute of Pathology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany.

出版信息

PLoS One. 2016 Nov 1;11(11):e0165181. doi: 10.1371/journal.pone.0165181. eCollection 2016.

DOI:10.1371/journal.pone.0165181
PMID:27802291
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5089548/
Abstract

BACKGROUND

Neuroendocrine lung cancer (NELC) represents 25% of all lung cancer cases and large patient collectives exist as formalin-fixed, paraffin-embedded (FFPE) tissue only. FFPE is controversially discussed as source for molecular biological analyses and reference genes for NELC are poorly establishes.

MATERIAL AND METHODS

Forty-three representative FFPE-specimens were used for mRNA expression analysis using the digital nCounter technology (NanoString). Based on recent literature, a total of 91 mRNA targets were investigated as potential tumor markers or reference genes. The geNorm, NormFinder algorithms and coefficient of correlation were used to identify the most stable reference genes. Statistical analysis was performed by using the R programming environment (version 3.1.1).

RESULTS

RNA integrity (RIN) ranged from 1.8 to 2.6 and concentrations from 34 to 2,109 ng/μl. However, the nCounter technology gave evaluable results for all samples tested. ACTB, CDKN1B, GAPDH, GRB2, RHOA and SDCBP were identified as constantly expressed genes with high stability (M-)values according to geNorm, NormFinder and coefficients of correlation.

CONCLUSION

FFPE-derived mRNA is suitable for molecular biological investigations via the nCounter technology, although it is highly degraded. ACTB, CDKN1B, GAPDH, GRB2, RHOA and SDCBP are potent reference genes in neuroendocrine tumors of the lung.

摘要

背景

神经内分泌肺癌（NELC）占所有肺癌病例的25%，且大量患者群体仅存在福尔马林固定、石蜡包埋（FFPE）组织。FFPE作为分子生物学分析的来源存在争议，且NELC的参考基因尚未很好地确立。

材料与方法

使用数字nCounter技术（NanoString）对43个代表性FFPE样本进行mRNA表达分析。基于近期文献，共研究了91个mRNA靶点作为潜在的肿瘤标志物或参考基因。使用geNorm、NormFinder算法和相关系数来鉴定最稳定的参考基因。使用R编程环境（版本3.1.1）进行统计分析。

结果

RNA完整性（RIN）范围为1.8至2.6，浓度为34至2109 ng/μl。然而，nCounter技术对所有测试样本均给出了可评估的结果。根据geNorm、NormFinder和相关系数，ACTB、CDKN1B、GAPDH、GRB2、RHOA和SDCBP被鉴定为具有高稳定性（M-）值的持续表达基因。

结论

尽管FFPE衍生的mRNA高度降解，但通过nCounter技术适用于分子生物学研究。ACTB、CDKN1B、GAPDH、GRB2、RHOA和SDCBP是肺神经内分泌肿瘤中有效的参考基因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ce5/5089548/5373ce5d1a17/pone.0165181.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ce5/5089548/3478a82422a4/pone.0165181.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ce5/5089548/5373ce5d1a17/pone.0165181.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ce5/5089548/3478a82422a4/pone.0165181.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ce5/5089548/5373ce5d1a17/pone.0165181.g002.jpg

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