Figure 5.

Effects of Aurora A/PLK1–associated long noncoding RNAs (APAL overexpression on cell cycle progression. A–B) The effect of APAL overexpression on tumor cell proliferation. APAL overexpression in MDA-MB-231 was detected by northern blot (A). CellTiter-Glo assay showed proliferation of MDA-MB-231 cells stably expressing APAL or control Sno-vector at a density of 1000 cells per well in 96-well plates. P value was based on the general linear model of repeated measures and was two-sided. C–D) Colony formation of MDA-MB-231 cells stably expressing APAL or control Sno-vector. A total of 1000 cells per well were seeded in six-well plates in triplicates. E) M-phase progression of APAL-overexpressing or control cells. MDA-MB-231 cells stably transfected with APAL or control vector were treated with nocodazole (100 ng/mL) for 16 hours. Mitotic cells were harvested using the mitotic shake-off method. Cell cycle profile was analyzed at the indicated time points after the cells were released. F) Mitosis progression of APAL-overexpressing or control MDA-MB-231 cells. G–J) Effects of chemotherapy drug treatments and anoikis on APAL-overexpressing or control MDA-MB-231 cells. Cells were treated with 5 μM cisplatin (48 hours), 5 nM docetaxel (24 hours), 200 nM cyclophosphamide (48 hours), or cultured in suspension (36 hours) followed by costaining of annexin V and propidium iodide (PI) for flow cytometric analysis. Graphs represent means ±SD of experimental triplicates. Two-sided independent sample t test was used to compare APAL-overexpressing and control cells in bar graphs.