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是人类肝癌组织和血液样本中RT-qPCR最适合的内参基因。

is the most suitable reference gene for RT-qPCR in human HCC tissues and blood samples.

作者信息

Ahn Hye Ri, Baek Geum Ok, Yoon Moon Gyeong, Son Ju A, You Donglim, Yoon Jung Hwan, Cho Hyo Jung, Kim Soon Sun, Cheong Jae Yeon, Eun Jung Woo

机构信息

Department of Gastroenterology, Ajou University School of Medicine, Suwon 16499, Republic of Korea.

Department of Biomedical Sciences, Ajou University Graduate School of Medicine, Suwon 16499, Republic of Korea.

出版信息

Oncol Lett. 2021 Nov;22(5):791. doi: 10.3892/ol.2021.13052. Epub 2021 Sep 17.

DOI:10.3892/ol.2021.13052
PMID:34584568
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8461756/
Abstract

Reverse transcription-quantitative (RT-q) PCR is the most feasible and useful technique for identifying and evaluating cancer biomarkers; however, the method requires suitable reference genes for gene expression analysis. The aim of the present study was to identify the most suitable reference gene for the normalization of relative gene expression in human hepatocellular carcinoma (HCC) tissue and blood samples. First, 14 candidate reference genes were selected through a systematic literature search. The expression levels of these genes ( and ) were evaluated using human multistage HCC transcriptome data (dataset GSE114564), which included normal liver (n=15), chronic hepatitis (n=20), liver cirrhosis (n=10), and early (n=18) and advanced HCC (n=45). From the 14 selected genes, five genes, whose expression levels were stable in all liver disease statuses ( and ), were further assessed using RT-qPCR in 40 tissues (20 paired healthy tissues and 20 tissues from patients with HCC) and 40 blood samples (20 healthy controls and 20 samples from patients with HCC). BestKeeper statistical algorithms were used to identify the most stable reference genes, of which was found to be the most stable in both HCC tissues and blood samples. Therefore, the results of the present study suggest as a promising reference gene for the normalization of relative RT-qPCR techniques in HCC-related studies.

摘要

逆转录定量（RT-q）PCR是鉴定和评估癌症生物标志物最可行且有用的技术；然而，该方法需要合适的内参基因用于基因表达分析。本研究的目的是确定用于人肝细胞癌（HCC）组织和血液样本中相对基因表达标准化的最合适内参基因。首先，通过系统的文献检索选择了14个候选内参基因。使用人类多阶段HCC转录组数据（数据集GSE114564）评估这些基因的表达水平，该数据集包括正常肝脏（n = 15）、慢性肝炎（n = 20）、肝硬化（n = 10）以及早期（n = 18）和晚期HCC（n = 45）。从14个选定的基因中，选择了5个在所有肝病状态下表达水平均稳定的基因，进一步在40个组织（20对健康组织和20个HCC患者组织）和40个血液样本（20个健康对照和20个HCC患者样本）中使用RT-qPCR进行评估。使用BestKeeper统计算法鉴定最稳定的内参基因，发现该基因在HCC组织和血液样本中均最稳定。因此，本研究结果表明该基因有望作为HCC相关研究中相对RT-qPCR技术标准化的内参基因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5fcc/8461756/41f17c2160fb/ol-22-05-13052-g02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5fcc/8461756/4dd925394b1a/ol-22-05-13052-g00.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5fcc/8461756/aa5ced7bf860/ol-22-05-13052-g01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5fcc/8461756/41f17c2160fb/ol-22-05-13052-g02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5fcc/8461756/4dd925394b1a/ol-22-05-13052-g00.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5fcc/8461756/aa5ced7bf860/ol-22-05-13052-g01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5fcc/8461756/41f17c2160fb/ol-22-05-13052-g02.jpg

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