Pre-clinical Characterization Of 4SC-202, A Novel Class ? HDAC Inhibitor,Against Colorectal Cancer Cells

Objective:In the current study, deacetylase (HDAC) inhibitor 4SC-202 in vivo effect of anti-colon cancer cells,a novel class I Histone deacetylase (HDAC) inhibitor (HDACi), in preclinical colon cancer models.Methods:1) MTT assay, trypan blue staining assay and colony formation assay were performed to test the potential role of 4SC-202 on colon cancer cell survival, proliferation and death;2) Histone DNA ELISA assay, Caspase-3/-9 activity assay, Annexin V-PI FACS assay and Western Blotting assay were performed to test colon cancer cell apoptosis following 4SC-202 treatment.3) PI-FACS assay was performed to examine cell cycle progression in 4SC-202-treated colon cancer cells.4) Pharmacological and shRNA strategies were applied to inhibit AKT activation in colon cancer cells,4SC-202-medaited in vitro anti-cancer activity in those colon cancer cells was tested.5) The impact MTT assay, Histone DNA ELISA method to detect relatively oxaliplatin group,4SC-202 group on survival apoptosis of colon group and oxaliplatin combined with and cancer cells6) HT-29 xenograft nude mice model was applied, the in vivo anti-colon cancer cell activity by 4SC-202 or together with oxaliplatin was analyzed.RESULTS:1)4SC-202 significantly inhibited colon cancer cells (primary human colon cancer cells and HT-29, HCT-116, HT-15, DLD-1 cell line) survival and proliferation inhibition in a time and concentration-dependent manner.2) 4SC-202 induced apoptosis in colon cancer3) 4SC-202-induced cytotoxicity was dramatically potentiated with serum starvation, AKT inhibition (by perifosine or MK-2206), or AKT1-shRNA knockdown in colon cancer cells. On the other hand, G1 and S phases colon cancer cells gradually reduce the proportion of increase in the proportion of G2-M phase.4) 4SC-202, at a low concentration, enhanced oxaliplatin-induced in vitro anti-colon cancer activity.5) Oral gavage of 4SC-202 inhibited HT-29 xenograft growth in nude mice, and when combined with oxaliplatin, its activity was further strengthened.Conclusions:1) 4SC-202 concentration and time-dependent inhibition of human colon cancer cell survival, proliferation and induce apoptosis in cancer cells;2) 4SC-202 colon cancer induced G2-M cell cycle arrest;3) AKT inhibition potentiates 4SC-202's activity in colon cancer cells4) 4SC-202 sensitizes the activity of oxaliplatin against colon cancer cells5) 4SC-202 and oxaliplatin synergistically inhibits HT-29 xenograft growth in nude mice...