Anti-tumor Effects And Molecular Mechanisms Of WYX56, A Novel Naphthalimide Derivant

Objective:Tumor cells and normal cells are different in DNA and the difference can play a vital role as targeting therapy. Therefore, DNA-damaging agents have been widely used in anticancer therapy. Naphthalimides can easily embed in DNA, and it has become one of the interesting field of investigation. In this paper, we investigated the anticancer effect of WYX56, a novel naphthalimide derivant, and explored the corresponding molecular mechanisms.Methods:The antiproliferative effects of WYX56 were assessed by MTT assay in Hep G2 and SMMC-7721 cells. Morphological change was observed by the inverted biological microscope. Apoptosis was assessed with AO/EB/Hoechst33342 fluorescent staining by HCS. Cell cycle of Hep G2 and SMMC-7721 was measured by Flow Cytometry with PI staining. Wound scratch and Transwell experiment were used to investigate migration and invasion. The expression of cyclinB1, CDK1 and other relative proteins were assessed by Western Blotting. Moreover, we study the antitumor effects of WYX56 in graft-tumor model.Results:The present data demonstrated that WYX56 inhibited growth effectively in Hep G2 cells and SMMC-7721 cells in dose- and time-dependent manner. The IC50(48 h) is(9.33±0.66) μM,(6.03±1.26) μM, respectively. Inverted biological microscope and AO/EB/Hoechst 33342 fluorescent staining both showed that WYX56 could induce the two cells cycle to be arrested. Flow cytometry testing confirmed that most of the cells were stuck in the G2/M phase and this phenomenon may be related to over-expression of p21. Through the wound scratch and Transwell experiment, WYX56 could inhibit the two cells migration and invasion in dose-dependent manner. Western blotting showed that WYX56 up-regulated expression of E–cadherin and down-regulated expression of Integrinα6. In-vivo experiment showed that WYX56 was highly anti-tumor active. The results demonstrated that WYX56 inhibited H22 tumor growth(Tumor-inhibition rate=52.63%) and interrupted lung metastasis and invasion(Inhibition rate=75.73%) in Swiss mice. In BLAB/c mice, WYX56 not only interrupted lung metastasis and invasion(Inhibition rate=60.66%) but also extended survival time. Furthermore, WYX56 could also interrupt lung metastasis in BLAB/c mice inoculated with CT-26 cells(Inhibition rate=70.89%).Conclusions:WYX56 could inhibit proliferation, migration and invasion of human liver cancer cells significantly in vitro and in vivo.