Selection of reference genes suitable for normalization of RT-qPCR data in glioma stem cells

被引：8

作者：

Dang, Weiqi ^{[1
,2
,3
]}

Zhang, Xiang ^{[1
,2
,3
]}

Ma, Qinghua ^{[1
,2
,3
]}

Chen, Lu ^{[1
,2
,3
]}

Cao, Mianfu ^{[1
,2
,3
]}

Miao, Jingya ^{[1
,2
,3
]}

Cui, Youhong ^{[1
,2
,3
]}

Zhang, Xia ^{[1
,2
,3
]}

机构：

[1] Army Med Univ, Mil Med Univ 3, Southwest Hosp, Inst Pathol, Chongqing 400038, Peoples R China

[2] Army Med Univ, Mil Med Univ 3, Southwest Hosp, Southwest Canc Ctr, Chongqing 400038, Peoples R China

[3] Minist Educ China, Key Lab Tumor Immunopathol, Chongqing 400038, Peoples R China

来源：

BIOTECHNIQUES | 2020年 / 68卷 / 03期

基金：

中国国家自然科学基金;

关键词：

glioma stem cells; quantitative real-time PCR; reference genes; stemness-related genes; tumor-initiating cells; REAL-TIME PCR; IDENTIFICATION; EXPRESSION;

D O I：

10.2144/btn-2019-0098

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Considering the importance of gene expression studies for understanding the biology of glioma stem cells (GSCs), we aimed to identify the reliable reference genes in GSCs that were derived from the glioma cell lines T98G, LN229, 090116 and 091214. Quantitative real-time reverse-transcription PCR was employed using 11 reference genes identified through a PubMed literature search, and the assessment of stability through the geNorm, Normfinder and coefficient of variation methods was performed to select suitable reference genes. We found that HPRT1 and RPL13A were the most suitable reference genes, and validated the geometric mean of these genes to normalize the expression of stemness genes by GSCs. Therefore, it is necessary to select novel cell-specific reference genes with greater expression stability for gene expression studies in GSCs. METHOD SUMMARY We employed the geNorm, Normfinder and coefficient of variation methods to identify novel reference genes, with greater expression stability in glioma stem cells, for the study of gene expression.

引用

页码：130 / 137

页数：8

共 31 条

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