PIK3CA and KRAS mutations in cell free circulating DNA are useful markers for monitoring ovarian clear cell carcinoma
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Asuka Morikawa1,2, Tomoatsu Hayashi1, Naomi Shimizu1, Mana Kobayashi1, Kenzui Taniue1, Akiko Takahashi1, Kota Tachibana1, Misato Saito2, Ayako Kawabata1,2, Yasushi Iida2, Kazu Ueda2, Motoaki Saito2, Nozomu Yanaihara2, Hiroshi Tanabe3, Kyosuke Yamada2, Hirokuni Takano3, Osamu Nureki4, Aikou Okamoto2 and Tetsu Akiyama1
1Laboratory of Molecular and Genetic Information, Institute of Molecular and Cellular Biosciences, The University of Tokyo, Tokyo, Japan
2Department of Obstetrics and Gynecology, Jikei University School of Medicine, Tokyo, Japan
3Department of Obstetrics and Gynecology, Jikei University, Kashiwa Hospital, Chiba, Japan
4Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan
Tetsu Akiyama, email: [email protected]
Keywords: OCCC; cfDNA; PIK3CA; KRAS; digital PCR
Received: June 16, 2017 Accepted: February 10, 2018 Epub: February 22, 2018 Published: March 16, 2018
Ovarian clear cell carcinoma (OCCC) exhibits distinct phenotypes, such as resistance to chemotherapy, poor prognosis and an association with endometriosis. Biomarkers and imaging techniques currently in use are not sufficient for reliable diagnosis of this tumor or prediction of therapeutic response. It has recently been reported that analysis of somatic mutations in cell-free circulating DNA (cfDNA) released from tumor tissues can be useful for tumor diagnosis. In the present study, we attempted to detect mutations in PIK3CA and KRAS in cfDNA from OCCC patients using droplet digital PCR (ddPCR). Here we show that we were able to specifically detect PIK3CA-H1047R and KRAS-G12D in cfDNA from OCCC patients and monitor their response to therapy. Furthermore, we found that by cleaving wild-type PIK3CA using the CRISPR/Cas9 system, we were able to improve the sensitivity of the ddPCR method and detect cfDNA harboring PIK3CA-H1047R. Our results suggest that detection of mutations in cfDNA by ddPCR would be useful for the diagnosis of OCCC, and for predicting its recurrence.
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