Identification of TMEM208 and PQLC2 as reference genes for normalizing mRNA expression in colorectal cancer treated with aspirin.
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.
Department of Gastrointestinal Medical Oncology, The Affiliated Tumor Hospital of Harbin Medical University, Harbin, China.
Yuanyuan
Y
Zhu
Department of Pathology, Harbin Medical University, Harbin, China.
Chao
C
Yang
Department of Pathology, Harbin Medical University, Harbin, China.
Mingjiao
M
Weng
Department of Nutrition and Food Hygiene, Public Health College, Harbin Medical University, Harbin, China.
Yan
Y
Zhang
Department of Sports Humanities, Harbin Sport University, Harbin, China.
Chunhui
C
Yang
Department of Pathology, Harbin Medical University, Harbin, China.
Yinji
Y
Jin
Department of Pathology, Harbin Medical University, Harbin, China.
Weiwei
W
Yang
Department of Pathology, Harbin Medical University, Harbin, China.
Yan
Y
He
Department of Pathology, Harbin Medical University, Harbin, China.
Yiqi
Y
Wu
Department of Pathology, Harbin Medical University, Harbin, China.
Yuhua
Y
Zhang
Department of Gastrointestinal Medical Oncology, The Affiliated Tumor Hospital of Harbin Medical University, Harbin, China.
Guangyu
G
Wang
Department of Pathology, Harbin Medical University, Harbin, China.
Riju James
RJ
RajkumarEzakiel Redpath
Department of Pathology, Harbin Medical University, Harbin, China.
Lei
L
Zhang
Department of Pathology, Harbin Medical University, Harbin, China.
Xiaoming
X
Jin
Department of Nutrition and Food Hygiene, Public Health College, Harbin Medical University, Harbin, China.
Ying
Y
Liu
Center of Digital Dentistry, Pecking University School and Hospital of Stomatology, Beijing, China.
Yuchun
Y
Sun
Department of Gastrointestinal Surgery, International Hospital of Pecking University, Beijing, China.
Ning
N
Ning
Department of Histology and Embryology, Harbin Medical University, Harbin, China.
Yu
Y
Qiao
Department of Microbiology, Harbin Medical University, Harbin, China.
Fengmin
F
Zhang
Department of Gastrointestinal Medical Oncology, The Affiliated Tumor Hospital of Harbin Medical University, Harbin, China.
Zhiwei
Z
Li
Department of Pathology, Harbin Medical University, Harbin, China.
Tianzhen
T
Wang
Department of Gastrointestinal Medical Oncology, The Affiliated Tumor Hospital of Harbin Medical University, Harbin, China.
Yanqiao
Y
Zhang
Department of Pathology, Harbin Medical University, Harbin, China.|The Northern Medicine Translational Center, Heilongjiang Province Academy of Medical Science, Harbin, China.
Xiaobo
X
Li
D000199:Actins / Q000235:genetics / Q000378:metabolism; D026923:Amino Acid Transport Systems, Basic / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D001241:Aspirin / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects; D015179:Colorectal Neoplasms / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology; D020869:Gene Expression Profiling / Q000592:standards*; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014404:Tubulin / Q000235:genetics / Q000378:metabolism; D014407:Tumor Cells, Cultured; D023041:Xenograft Model Antitumor Assays
D000199:Actins; D026923:Amino Acid Transport Systems, Basic; D000894:Anti-Inflammatory Agents, Non-Steroidal; D008565:Membrane Proteins; D012333:RNA, Messenger; C573925:SLC66A1 protein, human; C585265:TMEM208 protein, human; D014404:Tubulin; D001241:Aspirin
aspirin; colorectal cancer; reference gene