Enhanced expression of RAD51 associating protein-1 is involved in the growth of intrahepatic cholangiocarcinoma cells.
OBJECTIVE
Intrahepatic cholangiocarcinoma (ICC) is the second most common primary cancer in the liver, and its incidence is increasing in developed countries.


METHODS
To discover novel molecular targets for the diagnosis and treatment of ICCs, we earlier analyzed expression profiles of 25 ICCs using a cDNA microarray containing 27,648 genes. In this study, we focused on the RAD51 associating protein-1 (RAD51AP1) gene because its expression was frequently elevated in our microarray data.


RESULTS
Quantitative PCR confirmed that RAD51AP1 expression was elevated in the great majority of the ICCs examined. Immunohistochemical analysis with anti-RAD51AP1 antibody further corroborated its accumulation in 14 of 23 ICC tissues (61%). Notably, suppression of RAD51AP1 by short interfering RNA resulted in growth suppression of cholangiocarcinoma cells, suggesting its involvement in the development and/or progression of ICC. Because RAD51AP1 interacts with RAD51, a molecule involved in DNA repair, we investigated whether RAD51AP1 is implicated in DNA strand breaks using gamma-irradiation. As a result, gamma-irradiation augmented RAD51AP1 protein expression and brought a focus formation in the nuclei, where accumulated RAD51AP1 colocalized with phosphorylated histone 2AX (gamma-H2AX) and RAD51. These data suggest that RAD51AP1 may play a role in cell proliferation as well as DNA repair.


CONCLUSIONS
Our findings may contribute to the better understanding of cholangiocarcinogenesis and open a new avenue to the development of novel therapeutic and/or diagnostic approach to this type of tumor.
Obama
Kazutaka
K
Laboratory of Molecular Medicine, Human Genome Center and Division of Clinical Genome Research, Institute of Medical Science, University of Tokyo, Tokyo, Japan.
Satoh
Seiji
S
Hamamoto
Ryuji
R
Sakai
Yoshiharu
Y
Nakamura
Yusuke
Y
Furukawa
Yoichi
Y
D000818:Animals; D001650:Bile Duct Neoplasms / Q000235:genetics* / Q000473:pathology*; D001653:Bile Ducts, Intrahepatic / Q000473:pathology*; D015152:Blotting, Northern; D015153:Blotting, Western; D002454:Cell Differentiation; D049109:Cell Proliferation; D018281:Cholangiocarcinoma / Q000235:genetics* / Q000473:pathology*; D017931:DNA Primers / Q000737:chemistry; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000235:genetics*; D005720:Gamma Rays; D020869:Gene Expression Profiling; D006801:Humans; D007124:Immunoenzyme Techniques; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured / Q000528:radiation effects
D017931:DNA Primers; D004268:DNA-Binding Proteins; C110410:RAD51AP1 protein, human; D012333:RNA, Messenger; D016601:RNA-Binding Proteins