Fig. 5

miR-141 promotes ovarian cancer cell growth and tumor seeding by enhancing anoikis resistance of ovarian cancer cells. a Experimental overview of anoikis assay. b The relative apoptosis rates of miR-141 stable clones and KLF12 knockdown cells were measured using TUNEL assay. Green signals represent DAPI positive cells and red signals represent TUNEL positive cells. Both overexpression of miR-141 or knockdown of KLF12 resulted in lower apoptotic rates of ovarian cancer cells after treated by anoikis assay. c Flow cytometry cell cycle analysis revealed that percentages of sub-G1 phase in either KLF12 knockdown OVCA433 cells or miR-141 overexpressed SKOV3 cells were reduced as compared with their negative controls. d Western blotting showed that either knockdown of KLF12 or overexpression of miR-141 could suppress ovarian cancer cell apoptosis through reduction of cleaved-PARP and cleaved-caspase 3 expression in anoikis treated ovarian cancer cells (Upper: KLF12 knockdown clones in OVCA433, Lower; miR-141 stable expressing clones in SKOV3). Bar, 50 μM. e Effects of miR-141 on ovarian cancer growth and tumor seeding were revealed using in vivo model by i.p. injecting miR-141 and the scrambled control clones into 5-weeks old female nude mice. Average number of tumor nodules (Left) and the tumor burden (Right) formed by miR-141 clones were much higher than the scrambled control and they were observed across the peritoneal cavity as photographed on Day 28 (4 weeks). The white arrows indicate the location of tumor nodules