You have accessJournal of UrologyProstate Cancer: Basic Research & Pathophysiology II1 Apr 2016MP66-01 GENE KNOCKOUT OF NANOG AND NANOGP8 MEDIATED BY CRISPR/CAS9 DECREASES THE MALIGNANT POTENTIAL OF PROSTATE CANCER CELLS. Norihiko Kawamura, Keisuke Nimura, Hiromichi Nagano, Takahiro Yoshida, Atsunari Kawashima, Takeshi Ujike, Akira Nagahara, Kazutoshi Fujita, Motohide Uemura, Yasufumi Kaneda, and Norio Nonomura Norihiko KawamuraNorihiko Kawamura More articles by this author , Keisuke NimuraKeisuke Nimura More articles by this author , Hiromichi NaganoHiromichi Nagano More articles by this author , Takahiro YoshidaTakahiro Yoshida More articles by this author , Atsunari KawashimaAtsunari Kawashima More articles by this author , Takeshi UjikeTakeshi Ujike More articles by this author , Akira NagaharaAkira Nagahara More articles by this author , Kazutoshi FujitaKazutoshi Fujita More articles by this author , Motohide UemuraMotohide Uemura More articles by this author , Yasufumi KanedaYasufumi Kaneda More articles by this author , and Norio NonomuraNorio Nonomura More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.1275AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES NANOG is an essential transcription factor for self-renewal and pluripotency of embryonal stem cells. Recently, it has been reported that NANOG is expressed in various somatic cancers, including prostate cancer, and drive tumor development, and that increased NANOG expression in human prostate cancer tissues is correlated with an increased Gleason score. NANOG (hereinafter NANOG1) has many pseudogenes, and only NANOGP8 pseudogene encodes the full-length NANOG1 protein with high sequence similarity, and NANOGP8 is reported to be expressed in most of various cancer cells as a primary contributor of NANOG mRNA expression and to increase the malignant potential. However, the proportion of NANOG protein expression that comes from NANOG1 and NANOG8 in cancer cells is not known because of the high similarity between them. Therefore, a causal role of NANOG1 and NANOGP8 in prostate cancer cells is not clear. METHODS We established NANOG1-/- and NANOGP8-/- prostate cancer cell lines from DU145 cells using CRISPR/Cas9, and also we established NANOG1-rescued cells and NANOGP8-rescued cells from each NANOG knockout cells for rescue experiments. We examined cancer properties associated with malignant potential in these cells and DU145 cells, including self-renewal, sphere-formation, migration, drug resistance and tumorigenic potential. RESULTS Colony formation assays were performed to examine the role of NANOG1 and NANOGP8 in self-renewal. The colony-forming capacity of NANOG1-/- and NANOGP8-/- cells was decreased compared to parental cells. The sphere-forming capacity of NANOG1-/- and NANOGP8-/- cells decreased to approximately 50% compared to DU145 cells. Wound-healing assays were performed to examine the migration capacity, and migration was decreased in NANOG1-/- and NANOGP8-/- cells by 40-60%. MTS assays 48 hours after docetaxel administration were performed to evaluate the effect of NANOG1 and NANOGP8 on drug sensitivity. NANOG1-/- and NANOP8-/- cells showed increased sensitivity to docetaxel. NANOG1 and NANOGP8 knockout did not inhibit in vitro cell proliferation, but in vivo tumorigenic potential decreased significantly. These phenotypes were recovered in NANOG1- and NANOGP8-rescued cell lines. CONCLUSIONS These results indicate that both NANOG1 and NANOGP8 proteins are expressed in prostate cancer cell lines, and both genes equally contribute to the high malignant potential of prostate cancer cells. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e872 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Norihiko Kawamura More articles by this author Keisuke Nimura More articles by this author Hiromichi Nagano More articles by this author Takahiro Yoshida More articles by this author Atsunari Kawashima More articles by this author Takeshi Ujike More articles by this author Akira Nagahara More articles by this author Kazutoshi Fujita More articles by this author Motohide Uemura More articles by this author Yasufumi Kaneda More articles by this author Norio Nonomura More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...