In vitro Investigation of Anticancer, Cell-Cycle-Inhibitory, and Apoptosis-Inducing Effects of Diversin, a Natural Prenylated Coumarin, on Bladder Carcinoma Cells

Abstract

Chemotherapy is one of the main strategies for reducing the rate of cancer progression or, in some<br><br>cases, curing the tumour. Since a great number of chemotherapeutic agents are cytotoxic compounds,<br><br>i.e.similarly affect normal and neoplastic cells, application of antitumour drugs is preferred in cancer<br><br>management and therapy. In this study, the cytotoxicity of diversin was evaluated in 5637 cells, a tran-<br><br>sitional cell carcinoma (TCC) subline (bladder carcinoma), and normal human fibroblast cells using<br><br>the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2 H-tetrazolium bromide (MTT) assay. Chromatin con-densation and DNA damage induced by diversin were also determined by means of 40,6-diamidino-2-phenylindole (DAPI) staining and the comet assay, respectively. In addition, the mechanism of action<br><br>of diversin was studied in more detail by the caspase 3 colourimetric assay and flow cytometry-based<br><br>cell-cycle analyses (PI staining). Our results revealed that diversin has considerable cytotoxic effects<br><br>in 5637 cells, but not on HFF3 (human foreskin fibroblast) and HDF1 (human dermal fibroblast) cells.<br><br>Further studies showed that diversin exerts its cytotoxicity via induction of chromatin condensation,<br><br>DNA damage, and activation of caspase 3 in 5637 cells. In addition, flow cytometric analyses revealed<br><br>that 5637 cells are mostly arrested at the G2 phase of the cell cycle in the presence of diversin