RNA mutations of prox1 detected in human esophageal cancer cells by the shifted termination assay.
We have recently reported a novel finding that a candidate tumor suppressor gene prox1 suffered adenosine-to-inosine (A-to-I) RNA mutation without genomic mutation in a subset of human cancer cells and lost its function. Hence, screening of mutations in both cDNA and genomic DNA could be important in the analysis of causes for cancers. Here, we applied a sensitive, accurate, and simple method, called shifted termination assay (STA) for detection of an A-to-I RNA mutation (R334G) in prox1. We prepared PCR-amplified samples containing the target base of RNA mutation from cDNAs and genomic DNAs of various cell lines and clinical samples, to demonstrate that the STA method can be used to identify not only genomic mutations but also RNA mutations more effectively compared to sequencing. By means of STA, we found prox1 R334G RNA mutations but not genomic DNA mutations in 4 of 8 cases of esophageal cancers. This method can help us to detect RNA mutation effectively and progress research of a potential oncogenic principle.
Yoshimoto
Takanobu
T
Molecular & Cancer Research Unit, HMRO, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
Takahashi
Meiko
M
Nagayama
Satoshi
S
Watanabe
Go
G
Shimada
Yutaka
Y
Sakasi
Yoshiharu
Y
Kubo
Hajime
H
D002277:Carcinoma / Q000235:genetics; D045744:Cell Line, Tumor; D004252:DNA Mutational Analysis; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D004938:Esophageal Neoplasms / Q000378:metabolism*; D015972:Gene Expression Regulation, Neoplastic; D018398:Homeodomain Proteins / Q000096:biosynthesis* / Q000235:genetics*; D006801:Humans; D008957:Models, Genetic; D009154:Mutation; D012313:RNA / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D025521:Tumor Suppressor Proteins / Q000096:biosynthesis* / Q000235:genetics*; D000098551:Prospero-Related Homeobox 1 Protein
D017931:DNA Primers; D018076:DNA, Complementary; D018398:Homeodomain Proteins; D025521:Tumor Suppressor Proteins; D000098551:Prospero-Related Homeobox 1 Protein; D012313:RNA