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. 2022 Jan 28;12:748403. doi: 10.3389/fonc.2022.748403

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Copyright © 2022 Jiang, Siu, Wang, Leung, Chan, Cheung, Ngan and Chan

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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PFKFB3 promotes stemness in ovarian cancer. (A) Transwell migration/invasion assays of PFKFB3-suppressing tumorspheres and ALDH+CD44+ cells and PFKFB3-overexpressing ALDH-CD44- cells. Representative images of migrating tumorspheres with transient knockdown of PFKFB3 (left; scale bar, 100 μm). (B) Clonogenic assay of tumorspheres and ALDH+CD44+ cells transfected with control and siPFKFB3 duplexes cells. (C) Sphere formation assay of PFKFB3-suppressing tumorspheres and ALDH+CD44+ cells and PFKFB3-overexpressing ALDH-CD44- cells. Representative images of sphere-forming cells with transient knockdown of PFKFB3 (left; scale bar, 100 μm). (D) Relative KLF4 mRNA expression of PFKFB3-suppressing tumorspheres and ALDH+CD44+ cells and PFKFB3-overexpressing ALDH-CD44- cells, determined by qPCR. (E) Protein expression of PFKFB3, KLF4, BMI1, p-p65 and p65 in PFKFB3-knockdown tumorspheres (left) ALDH+CD44+ (middle) SKOV3 cells and PFKFB3-overexpressing ALDH-CD44- (right) SKOV3 cells, determined by immunoblotting. (F) After DMAPT (1 μM) treatment of PFKFB3-overexpressing ALDH-CD44- cells for 48 h, sphere formation assays were conducted and analyzed. (n = 3; *p < 0.05, **p < 0.01).