CRISPR/Cas9-mediated gene knockout of NANOG and NANOGP8 decreases the malignant potential of prostate cancer cells
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Norihiko Kawamura1,2, Keisuke Nimura1, Hiromichi Nagano1, Sohei Yamaguchi1, Norio Nonomura2, Yasufumi Kaneda1
1Division of Gene Therapy Science, Osaka University Graduate School of Medicine, Suita, Osaka 565–0871, Japan
2Department of Urology, Osaka University Graduate School of Medicine, Suita, Osaka 565–0871, Japan
Keisuke Nimura, e-mail: firstname.lastname@example.org
Yasufumi Kaneda, e-mail: email@example.com
Keywords: NANOG, NANOGP8, gene knockout, CRISPR/Cas9, prostate cancer
Received: March 31, 2015 Accepted: June 03, 2015 Published: June 15, 2015
NANOG expression in prostate cancer is highly correlated with cancer stem cell characteristics and resistance to androgen deprivation. However, it is not clear whether NANOG or its pseudogenes contribute to the malignant potential of cancer. We established NANOG- and NANOGP8-knockout DU145 prostate cancer cell lines using the CRISPR/Cas9 system. Knockouts of NANOG and NANOGP8 significantly attenuated malignant potential, including sphere formation, anchorage-independent growth, migration capability, and drug resistance, compared to parental DU145 cells. NANOG and NANOGP8 knockout did not inhibit in vitro cell proliferation, but in vivo tumorigenic potential decreased significantly. These phenotypes were recovered in NANOG- and NANOGP8-rescued cell lines. These results indicate that NANOG and NANOGP8 proteins are expressed in prostate cancer cell lines, and NANOG and NANOGP8 equally contribute to the high malignant potential of prostate cancer.
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