Identification of TMEM208 and PQLC2 as reference genes for normalizing mRNA expression in colorectal cancer treated with aspirin
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Yuanyuan Zhu1, Chao Yang2,Mingjiao Weng2, Yan Zhang3, Chunhui Yang4, Yinji Jin2,Weiwei Yang2, Yan He2, Yiqi Wu2, Yuhua Zhang2, Guangyu Wang1, Riju James RajkumarEzakiel Redpath2, Lei Zhang2, Xiaoming Jin2, Ying Liu3, Yuchun Sun5, Ning Ning6, Yu Qiao7, Fengmin Zhang8, Zhiwei Li1, Tianzhen Wang2, Yanqiao Zhang1, Xiaobo Li2,9
1Department of Gastrointestinal Medical Oncology, The Affiliated Tumor Hospital of Harbin Medical University, Harbin, China
2Department of Pathology, Harbin Medical University, Harbin, China
3Department of Nutrition and Food Hygiene, Public Health College, Harbin Medical University, Harbin, China
4Department of Sports Humanities, Harbin Sport University, Harbin, China
5Center of Digital Dentistry, Pecking University School and Hospital of Stomatology, Beijing, China
6Department of Gastrointestinal Surgery, International Hospital of Pecking University, Beijing, China
7Department of Histology and Embryology, Harbin Medical University, Harbin, China
8Department of Microbiology, Harbin Medical University, Harbin, China
9The Northern Medicine Translational Center, Heilongjiang Province Academy of Medical Science, Harbin, China
Yanqiao Zhang, email: firstname.lastname@example.org
Xiaobo Li, email: email@example.com
Tianzhen Wang, email: firstname.lastname@example.org
Keywords: aspirin, reference gene, colorectal cancer
Received: June 29, 2016 Accepted: January 23, 2017 Published: February 08, 2017
Numerous evidences indicate that aspirin usage causes a significant reduction in colorectal cancer. However, the molecular mechanisms about aspirin preventing colon cancer are largely unknown. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) is a most frequently used method to identify the target molecules regulated by certain compound. However, this method needs stable internal reference genes to analyze the expression change of the targets. In this study, the transcriptional stabilities of several traditional reference genes were evaluated in colon cancer cells treated with aspirin, and also, the suitable internal reference genes were screened by using a microarray and were further identified by using the geNorm and NormFinder softwares, and then were validated in more cell lines and xenografts. We have showed that three traditional internal reference genes, β-actin, GAPDH and α-tubulin, are not suitable for studying gene transcription in colon cancer cells treated with aspirin, and we have identified and validated TMEM208 and PQLC2 as the ideal internal reference genes for detecting the molecular targets of aspirin in colon cancer in vitro and in vivo. This study reveals stable internal reference genes for studying the target genes of aspirin in colon cancer, which will contribute to identify the molecular mechanism behind aspirin preventing colon cancer.
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