IR@PKUHSC  > 北京大学基础医学院  > 病原生物学系
学科主题基础医学
Selection of reference genes for RT-qPCR analysis in tumor tissues from male hepatocellular carcinoma patients with hepatitis B infection and cirrhosis
Liu, Shuang1; Zhu, Pengfei2,3; Zhang, Ling4; Ding, Shanlong2,3; Zheng, Sujun1; Wang, Yang1; Lu, Fengmin2,3
关键词Hepatocellular carcinoma hepatitis B virus liver cirrhosis male gender C-terminal binding protein 1 reference gene RT-qPCR
刊名CANCER BIOMARKERS
2013
DOI10.3233/CBM-130365
13期:5页:345-349
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Oncology
研究领域[WOS]Oncology
关键词[WOS]GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE GENE ; POLYMERASE-CHAIN-REACTION ; VIRUS-RNA ; EXPRESSION ; PCR ; PROTEIN ; CANCER
英文摘要

BACKGROUND: Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) has been widely used to quantify relative gene expression because of the high specificity, sensitivity and accuracy of this technique. However, its reliability is strongly depends on the expression stability of reference gene used for data normalization. Therefore, identification of reliable and condition specific reference genes is critical for the success of RT-qPCR.

OBJECTIVE: Hepatitis B virus (HBV) infection, male gender and the presence of cirrhosis are widely recognized as the leading independent risk factors for the development of hepatocellular carcinoma (HCC). This study aimed to select reliable reference gene for RT-qPCR analysis in HCC patients with all of those risk factors.

METHODS: Six candidate reference genes were analyzed in 33 paired tumor and non-tumor tissues from untreated HCC patients. The genes expression stabilities were assessed by geNorm and NormFinder.

RESULTS: C-terminal binding protein 1(CTBP1) was the most stable gene among the 6 candidate genes evaluated by both geNorm and NormFinder. The expression stability values were 0.08 for CTBP1 and UBC, 0.09 for HPRT1, 0.12 for HMBS, 0.14 for GAPDH and 0.18 for 18S with geNorm analysis. The stability values suggested by NormFinder software were CTBP1: 0.044, UBC: 0.063, HMBS: 0.072, HPRT1: 0.072, GAPDH: 0.098 and 18S rRNA: 0.161.

CONCLUSION: This is the first systematic analysis which suggested CTBP1 as the highest expression-stable gene in human male HBV infection related-HCC with cirrhosis. We recommend CTBP1 as the best candidate reference gene when RT-qPCR was used to determine gene(s) expression in HCC. This may facilitate the relevant HBV related HCC studies in the future.

语种英语
WOS记录号WOS:000329992500006
项目编号2012ZX10002005
资助机构Ministry of Science and Technology of the People&prime ; s Republic of China
引用统计
被引频次:9[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
版本出版稿
条目标识符http://ir.bjmu.edu.cn/handle/400002259/53185
专题北京大学基础医学院_病原生物学系
北京大学基础医学院
作者单位1.Henan Canc Hosp, Zhengzhou, Henan, Peoples R China
2.Capital Med Univ, Beijing Youan Hosp, Beijing Artificial Liver Treatment & Training Ctr, Beijing, Peoples R China
3.Peking Univ, Hlth Sci Ctr, Dept Microbiol, Beijing 100191, Peoples R China
4.Peking Univ, Hlth Sci Ctr, Ctr Infect Dis, Beijing 100191, Peoples R China
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GB/T 7714
Liu, Shuang,Zhu, Pengfei,Zhang, Ling,et al. Selection of reference genes for RT-qPCR analysis in tumor tissues from male hepatocellular carcinoma patients with hepatitis B infection and cirrhosis[J]. CANCER BIOMARKERS,2013,13(5):345-349.
APA Liu, Shuang.,Zhu, Pengfei.,Zhang, Ling.,Ding, Shanlong.,Zheng, Sujun.,...&Lu, Fengmin.(2013).Selection of reference genes for RT-qPCR analysis in tumor tissues from male hepatocellular carcinoma patients with hepatitis B infection and cirrhosis.CANCER BIOMARKERS,13(5),345-349.
MLA Liu, Shuang,et al."Selection of reference genes for RT-qPCR analysis in tumor tissues from male hepatocellular carcinoma patients with hepatitis B infection and cirrhosis".CANCER BIOMARKERS 13.5(2013):345-349.
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