ABSTRACT
TW-37 is a small molecule B cell lymphoma-2 (Bcl-2) homology 3 mimetic with potential anticancer activities. However, the in vivo anti-cancer effect of TW-37 in human oral cancer has not been properly studied yet. Here, we attempted to confirm antitumor activity of TW37 in human oral cancer. TW-37 significantly inhibited cell proliferation and increased the number of dead cells in MC-3 and HSC-3 human oral cancer cell lines. TW-37 enhanced apoptosis of both cell lines evidenced by annexin V/propidium iodide double staining, sub-G1 population analysis and the detection of cleaved poly (ADP-ribose) polymerase and caspase-3. In addition, TW-37 markedly downregulated the expression of Bcl-2 protein, while not affecting Bcl-xL or myeloid cell leukemia-1. In vivo, TW-37 inhibited tumor growth in a nude mice xenograft model without any significant liver and kidney toxicities. Collectively, these data reveal that TW-37 may be a promising small molecule to inhibit human oral cancer.
Subject(s)
Animals , Humans , Mice , Apoptosis , Caspase 3 , Cell Line , Cell Proliferation , Heterografts , Kidney , Liver , Mice, Nude , Mouth Neoplasms , Myeloid CellsABSTRACT
Objective To study the effect and mechanisms of TW-37 on cell proliferation,apoptosis,invasion and angiogenesis in pancreatic cancer cells in vitro and further explore the potential mechanism.Methods BxPC3 and HPAC cells were pretreated with TW-37 using untransfected or transfected with NF-κB p65 cDNA(p65 cDNA)or NF-κB p65 siRNA(siRNA-p65)cells as controls.Cell viability was determined by MrTT assay.Cell apoptosis was assessed by enzyme-linked immunosorbent assay (ELISA).Cell invasion and angiogenesis was detected by Transwell and endothelial tube formation assay of HUVECs.ELISA assay was used to measure the activity of NF-κB,and its target proteins of MMP-9 and VEGF were detected by western blot.Results TW-37 suppressed cell growth and induced apoptosis (A405:1.29 ± 0.21 vs 0.09 ± 0.01,1.07 s0.18 vs 0.08 ± 0.01),inhibited NF-κB activity and protein expression of NF-κB p65,VEGF and MMP-9(all P ＜0.05)in a dose-and time-dependent manner.The number of cells that invaded across the matrigel in the transwell chamber was (46.7 ±5.24) and (10.3 ± 1.26)/×200 in BxPC3 control and 0.75 μmol/L TW-37 group (P=0.001).The number of tube formation was (39.4 ±4.36) and (7.84 ± 1.25)/×200,(P =0.001).NF-κB activity was increased by p65 cDNA transfection,and decreased by TW-37 treatment in both of the two cell lines (P ＜0.05).However,NF-κB activity was decreased by p65 siRNA transfection,and greatly decreased by TW-37 treatment in both two cell lines (P ＜0.05 or P ＜0.01).Transfection of p65 cDNA did not significantly affect cell apoptosis.Transfection of p65 siRNA increased cell apoptosis,and greatly increased by TW-37 treatment in both two cell lines (all P ＜ 0.01).Conclusions TW-37 could inhibit the proliferation,invasion and angiogenesis in pancreatic cancer cells by regulating NF-κB signal pathway.