Combination treatment of ACY-241 and JQ1 synergistically inhibit HNSCC cell invasion and migration. (A,B) Wound healing assay of 2A3 and FaDu cells. Cells were seeded in 6-well plates and then scratched after cell adhesion. Artificial lines were drawn on microscopic images to visualize the scratched area. Percentage of wound closure was determined by the difference between wound areas in 0 h and 48 h. The wound was photographed at 50× magnification. Scale bar = 50 μm. (C) Transwell migration assay and (D) transwell invasion assay of 2A3 and FaDu cells. The cells were photographed at 200× magnification. Scale bar = 100 μm. Matrigel (0.4 mg/mL) was coated on the inner inserts of transwell plate for 1 h during invasion assay. Cells were treated with 0.2% DMSO, ACY-241 (4 μM), and JQ1 (2 μM) alone or in combination for 48 h. Data were normalized by cell viability to represent migration and invasion of viable cells. Values represent mean ± SD (n = 3). * p < 0.05, or *** p < 0.001 vs. DMSO control, $$$
p < 0.001 vs. ACY-241-treated group, #
p < 0.05, ##
p < 0.01 or ###
p < 0.001 vs. JQ1-treated group.