Figures & data
Figure 1 UNBS5162 and amonafide inhibits the proliferation of melanoma cells. (A and B) The proliferation of M14 or A375 cells treated with various concentrations of UNBS5162 and amonafide, respectively. (C) Cell-counting kit-8 assay was performed in order to estimate the proliferation of melanoma cells treated with 10 µM UNBS5162 or 8 µM amonafide; the OD value of the UNBS5162 and amonafide groups decreased significantly compared with the negative control (NC) group both in M14 and A375 cells. (D and E) Clone numbers of melanoma cells treated with 10 µM UNBS5162 or 8 µM amonafide decreased significantly compared with the NC group.
Notes: *P<0.05, the UNBS5162 group vs the NC group; #P<0.05, the amonafide group vs the NC group.
Figure 2 UNBS5162 and amonafide inhibits the migration and invasion of melanoma cells. (A and B) Representative images of the transwell assay for migration and invasion in melanoma cells with treatment of UNBS5162. (C and D) Representative images of transwell assay for migration and invasion in melanoma cells with treatment of amonafide. Invasion or migration cell numbers decreased significantly in UNBS5162 or amonafide treated cells when compared with negative control (NC) group both in M14 and A375 cells.
Note: *P<0.05.
Figure 3 UNBS5162 and amonafide promote the apoptosis of melanoma cells. (A) Cell apoptosis was detected by flow cytometry. (B) Apoptosis cell number of M14 and A375 cells treated with 10 µM UNBS5162 or 8 µM amonafide decreased significantly when compared with negative control (NC) cells. (C and E) The expressions of apoptotic-related genes were detected by Western blot. (D and F) Active Caspase3 and Bax expression were inhibited in UNBS5162 and amonafide groups, while Bcl-2 was promoted when compared with NC cells.
Note: *P<0.05.
Abbreviations: PI, propidium iodide; FITC, fluorescein isothiocyanate.
Figure 4 UNBS5162 and amonafide inhibit the activation of the AKT/mTOR pathway and reverse the epithelial–mesenchymal transition process in human melanoma cells. (A–D) Western blot was performed to confirm the pathway by which UNBS5162 and amonafide inhibited tumor progression; phosphorylation of AKT and mTOR were inhibited in M14 and A375 cells treated with 10 µM UNBS5162 or 8 µM amonafide for 48 hours, as well as P70/S6K and Cyclin D1. (C–H) The expressions of N-cad, Snail, Vimentin, and slug in UNBS5162 and amonafide groups decreased significantly, whereas E-cad increased significantly.
Note: *P<0.05.
Abbreviation: NC, negative control.
Figure 5 5FU combined with UNBS5162 or amonafide inhibits the proliferation of human melanoma cells. (A) CCK8 assay was performed to confirm the proliferation of cells when treated with 5FU combined with 10 µM UNBS5162. (B) CCK8 assay was performed to confirm the proliferation of cells with treatment of combined 5FU and 8 µM amonafide.
Notes: *P<0.05 vs the NC group; #P<0.05 vs the 5FU group.
Abbreviations: CCK8, cell-counting kit-8; NC, negative control
