3-Deoxysappanchalcone isolated from Caesalpinia sinensis shows anticancer effects on HeLa and PC3 cell lines: invasion, migration, cell cycle arrest, and signaling pathway

Abstract

To study the antitumor activity of compound 3-desoxysulforaphane (3-DSC) isolated from Caesalpinia sinensis, SRB assay, clone formation assay, flow cytometric cell cycle assay, scratch assay, transwell assay, and molecular docking were used to investigate the inhibitory effect of 3-DSC on HeLa and PC3 cells. The results showed that 3-DSC inhibited the cell migration and invasion by down-regulating expression of N-cadherin, Vimentin, MMP-2, and MMP-9 in HeLa and PC3 cells; It also inhibits cell proliferation by promoting the expression of CDK1 (cyclin-dependent kinases 1) and CDK2 (cyclin-dependent kinases 2), which arrests the tumor cell cycle at G2 phase. 3-DSC inhibits phosphorylation of AKT and ERK and upregulates the expression of the tumor suppressor gene p53. Molecular docking results confirmed that 3-DSC could bind firmly to AKT. In conclusion, 3-DSC inhibited the proliferation, migration and invasion of HeLa and PC3 cells.

Keywords: Caesalpinia sinensis; Cell cycle; Invasion; Migration; Proliferation.

Figure 1

Effect of 3-DSC on cell proliferation in HeLa and PC3 cells. A: Chemical structure of 3-DSC. B: SRB cell viability assay. C and D: Clone formation assay. ∗ indicates comparison with 3-DSC free group (0 group), ∗p < 0.05, ∗∗p < 0.01,∗∗∗p < 0.001.

Figure 2

Effect of 3-DSC on migration and invasio in HeLa and PC3 cells. A and B: wound Healing Assay. C and D: Transwell migration. E and F: Transwel invasion. G and H: The proteins expression of N-cadherin, Vimentin, MMP-2, and MMP-9. ∗ indicates comparison with 3-DSC free group (0 group), ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Figure 3

3-DSC effects on cell cycle. A and B: Cell cycle. C: Schematic representation of CDK1 and CDK2 regulation of cell cycle. D and E: The proteins expression of CDK1 and CDK2. ∗ indicates comparison with 3-DSC free group (0 group), ∗p < 0.05, ∗∗p < 0.01,∗∗∗p < 0.001.

Figure 4

Effect of 3-DSC on the mode and mechanism of death of HeLa and PC3. A: SRB cell viability assay to detect the mode of death. B and C: The proteins expression of AKT, p-ERK and p53. D and E: Molecular docking results of 3-DSC with AKT. Z-VAD-FMK is a Pan-Caspase inhibitor; MHY1485 is a potent, cell-permeable mTOR agonist that also effectively inhibits autophagy; Fer-1 (Ferrostatin-1) is a potent and selective inhibitor of ferroptosis; Nec-1 (Necrostatin-1) is a specific RIP1 (RIPK1) inhibitor that inhibits cell necrosis; Lip-1 (Liproxstatin-1) is a potent inhibitor of ferroptosis. ∗ indicates comparison with 3-DSC free group (0 group), ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.